CN100413861C - Compounds and methods for the treatment or prevention of flavivirus infections - Google Patents
Compounds and methods for the treatment or prevention of flavivirus infections Download PDFInfo
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- CN100413861C CN100413861C CNB2003801094491A CN200380109449A CN100413861C CN 100413861 C CN100413861 C CN 100413861C CN B2003801094491 A CNB2003801094491 A CN B2003801094491A CN 200380109449 A CN200380109449 A CN 200380109449A CN 100413861 C CN100413861 C CN 100413861C
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D333/00—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom
- C07D333/02—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings
- C07D333/04—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom
- C07D333/26—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D333/30—Hetero atoms other than halogen
- C07D333/36—Nitrogen atoms
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/38—Heterocyclic compounds having sulfur as a ring hetero atom
- A61K31/381—Heterocyclic compounds having sulfur as a ring hetero atom having five-membered rings
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/445—Non condensed piperidines, e.g. piperocaine
- A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
- A61K31/4535—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a heterocyclic ring having sulfur as a ring hetero atom, e.g. pizotifen
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/55—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- C07D—HETEROCYCLIC COMPOUNDS
- C07D333/00—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom
- C07D333/02—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings
- C07D333/04—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom
- C07D333/26—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D333/38—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
- C07D333/40—Thiophene-2-carboxylic acid
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- C07D—HETEROCYCLIC COMPOUNDS
- C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
- C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
- C07D409/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
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Abstract
The present invention provides novel compounds represented by formula (I) or pharmaceutically acceptable salts thereof useful for treating flaviviridae viral infection.
Description
Invention field
The present invention relates to compounds and use the method for this compounds for treating or prevention of flavivirus infections.
Background of invention
Hepatitis is the disease that all there is generation in a kind of whole world.Hepatitis generally is that virus causes, although other known reasons are also arranged.Viral hepatitis is modal hepatitis form at present.Have 750,000 Americans to be subjected to the influence of hepatitis every year approximately, wherein surpasses 150,000 people and be subjected to hepatitis C C virus (" HCV ") infection.
HCV be a kind of belong to flavivirus (Flavivirus) section just-strand rna virus, with pestivirus substantial connection is arranged, pestivirus comprises Pestivirus suis and bovine viral diarrhea virus (BVDV).It is believed that HCV by produce complementary negative-chain RNA template duplicates.Lack the effective shift copy system of this virus by hand, separate the HCV particle, show the about 50-60nm of its diameter with electron microscope from the human plasma that merges.The HCV genome is about 9, list-chain of 600bp, just-Yi RNA, and the amino acid whose polyprotein of coding 3009-3030, it cuts altogether by cell and two virus proteases and translation becomes sophisticated virus protein (core, E1, E2, p7, NS2, NS3, NS4A, NS4B, NS5A, NS5B albumen).It is believed that structural protein E1 and E2, most sugars albumen is embedded in and forms stable heterodimer in the viral lipid coating.Think that also the interaction of structural core protein and viral RNA genome forms nucleocapsid.The nonstructural proteins of called after NS2 to NS5 has the enzyme function of virus replication and protein processing, comprises polysaccharase, protease and helicase.
The main thing source that polluted by HCV is a blood.Popular explanation HCV gradient of infection among the high risk population has become a health problem.For example, in western countries, the hemophilia people of 60-90% and surpass 80% intravenous drug misuser all chronic infection HCV.To the intravenous pharmacy misuser, according to crowd's difference of being studied, prevalence rate is about 28% to 70%.The New Development HCV infection proportion that the blood transfusion back is relevant obviously descends owing to screen the improvement of blood donor's diagnostic method in recent years.
It is interferon-' alpha ' (1FN-α) that present HCV infects available unique only methods of treatment.Yet according to different clinical studyes, only patients serum's alanine aminotransferase (ALT) level of being treated of 70% transfers to normally, but after stopping IFN, these reactors have the 35-45% recurrence.Generally, only the patient of 20-25% to the long reaction that has of IFN.Clinical study shows: IFN and virazole (RIBA) combination therapy can produce more excellent clinical response than single with the IFN treatment.Treatment has differential responses to the different genotypes of HCV to IFN, and genotype 1b more tolerates the IFN treatment than 2 types and 3 types.
Therefore, be starved of the research and development anti-virus formulation.
Summary of the invention
On the one hand, the invention provides the compounds of class following formula representative, or its pharmacy acceptable salt:
In the formula;
Z is selected from: 3-7 unit's heterocycle or 3-7 unit cycloalkyl;
Y is a 6-10 unit aryl;
X is a 3-10 unit cycloalkyl;
M is the 0-1 integer;
Condition is, when Y is when not being substituted phenyl, X is not the 4-methylcyclohexane.
On the other hand, provide the method that flavivirus infects among a kind of treatment or the prevention host, comprise the The compounds of this invention, composition or the drug combination that give object treatment significant quantity.
On the other hand, a kind of combination that comprises The compounds of this invention and one or more other preparations is provided, described other preparations are selected from: virus serine protease inhibitors, viral polymerase inhibitors and viral helicase inhibitor, immunomodulator, antioxidant, antiseptic-germicide or antisense agents (antisense).
On the other hand, provide a kind of pharmaceutical composition, it comprises at least a The compounds of this invention and at least a pharmaceutically acceptable carrier or vehicle.
Another aspect provides The compounds of this invention, and composition or drug combination are treated or prevented host's flavivirus to infect.
Also have on the one hand, provide The compounds of this invention to be used for suppressing or reducing host's varial polymerases activity.
The medicine that provides The compounds of this invention to be used to make treatment or to prevent host's virus Flavivirus to infect is be provided on the one hand.
Detailed description of the invention
In the embodiment, The compounds of this invention comprises in the following embodiment those compounds of medication separately or drug combination.
In one embodiment of the present invention, Z is selected from 3-7 unit's heterocycle or 3-7 unit cycloalkyl.
In the embodiment, Z is:
In the formula;
W is CR
10R
11S (O) n, O or NR
12
Wherein, n is 0-2;
R
10And R
11Be H independently of one another, C
1-6Alkyl, C
6-10Aryl, C
3-10Heterocycle, C
3-10Heteroaralkyl, C
6-10Aralkyl, C (O)-C
1-6Alkyl, C
1-6Alkoxyl group, hydroxyl or formyl radical;
Or R
10And R
11Formation=O together ,=S or=N-Ra, wherein Ra is H, hydroxyl or C
1-6Alkyl; R
12Be H, C
1-6Alkyl, C
6-14Aryl, C
3-12Heterocycle, C
3-12Heteroaralkyl, C
6-16Aralkyl, C (O)-C
1-6Alkyl or C
1-6Alkoxyl group;
P is the integer of 1-3;
Q is the integer of 0-2;
R
13Be one or more optional substituting groups, they independently are selected from separately: halogen, nitro, nitroso-group, SO
3Rf, SO
2Rf, PO
3RcRd, CONRgRh, C
1-6Alkyl, C
2-6Thiazolinyl, C
2-6Alkynyl, C
6-12Aralkyl, C
6-12Aryl, C
1-6Alkoxyl group, C
2-6Alkene oxygen base, C
2-6Alkynyloxy group, C
6-12Aryloxy, C (O) C
1-6Alkyl, C (O) C
2-6Thiazolinyl, C (O) C
2-6Alkynyl, C (O) C
6-12Aryl, C (O) C
6-12Aralkyl, C
3-10Heterocycle, hydroxyl, NRgRh, C (O) ORf, cyano group, azido-, amidino groups or guanidine radicals; Rf wherein, Rc, Rd, Rg and Rh are H independently of one another, C
1-6Alkyl, C
2-6Thiazolinyl, C
2-6Alkynyl, C
6-10Aryl, C
3-10Heterocycle, C
3-10Heteroaralkyl or C
6-10Aralkyl;
Or Rc and Rd form a 5-10 unit heterocycle with oxygen;
Or Rg and Rh form a 3-10 unit heterocycle with nitrogen.
In the another embodiment, Z is 6-7 unit's heterocycle or 6-7 unit cycloalkyl.
In the embodiment, Z is a cyclohexyl, piperidyl or N (C
1-6Alkyl)-and piperidyl, azepanyl, methyl azepanyl, N (C
1-6Alkyl)-and piperidino methyl, THP trtrahydropyranyl, piperidino methyl, pyridyl, pyridylmethyl, tetrahydro thiapyran base, dioxolane ylmethyl or alkyl dioxin methyl, they are not substituted separately or are selected from following substituting group and replace by one or more: halogen, nitro, nitroso-group, SO
3Rf,
SO
2Rf, PO
3RcRd, CONRgRh, C
1-6Alkyl, C
2-6Thiazolinyl, C
2-6Alkynyl, C
6-12Aralkyl, C
6-12Aryl, C
1-6Alkoxyl group, C
2-6Alkene oxygen base, C
2-6Alkynyloxy group, C
6-12Aryloxy, C (O) C
1-6Alkyl, C (O) C
2-6Thiazolinyl, C (O) C
2-6Alkynyl, C (O) C
6-12Aryl, C (O) C
6-12Aralkyl, C (O) NHRf, C
3-10Heterocycle, hydroxyl, NRgRh, C (O) ORf, cyano group, azido-, amidino groups or guanidine radicals;
Rf wherein, Rc, Rd, Rg and Rh are H independently of one another, C
1-6Alkyl, C
2-6Thiazolinyl, C
2-6Alkynyl, C
6-10Aryl, C
3-10Heterocycle, C
3-10Heteroaralkyl or C
6-10Aralkyl;
Or Rc and Rd form a 5-10 unit heterocycle with oxygen;
Or Rg and Rh form a 3-10 unit heterocycle with nitrogen.
In the another embodiment, Z is a cyclohexyl, piperidyl, N (C
1-6Alkyl)-and piperidyl, azepanyl, methyl azepanyl, N (C
1-6Alkyl)-and piperidino methyl, THP trtrahydropyranyl, piperidino methyl, pyridyl, pyridylmethyl, tetrahydro thiapyran base, dioxolane ylmethyl or alkyl dioxin methyl, they are not substituted separately or are selected from following substituting group and replace by one or more: halogen, SO
2Rf, PO
3RcRd, CONRgRh, C
1-6Alkyl, C
6-12Aralkyl, C
6-12Aryl, C
1-6Alkoxyl group, C
6-12Aryloxy, C (O) C
1-6Alkyl, C (O) C
6-12Aryl, C (O) C
6-12Aralkyl, C (O) NHRf, C
3-10Heterocycle, hydroxyl, NRgRh, C (O) ORf, cyano group, azido-, amidino groups or guanidine radicals;
Wherein, Rf, Rc, Rd, Rg and Rh are H independently of one another, C
1-6Alkyl, C
2-6Thiazolinyl, C
2-6Alkynyl, C
6-12Aryl, C
3-10Heterocycle, C
3-10Heteroaralkyl or C
6-10Aralkyl.
In the embodiment, Z is a cyclohexyl, piperidyl, N (C
1-6Alkyl)-and piperidyl, azepanyl, methyl azepanyl, N (C
1-6Alkyl)-and piperidino methyl, THP trtrahydropyranyl, piperidino methyl, pyridyl, pyridylmethyl, tetrahydro thiapyran base, dioxolane ylmethyl or alkyl dioxin methyl are not substituted separately or are selected from following substituting group and replace by one or more: halogen, SO
2Rf, CONRgRh, C
1-6Alkyl, C
6-12Aralkyl, C
6-12Aryl, C
1-6Alkoxyl group, C (O) C
1-6Alkyl, C (O) NHRf, C
3-10Heterocycle, hydroxyl, NRgRh, C (O) Orf or cyano group;
Rf wherein, Rg and Rh are H or C independently of one another
1-6Alkyl.
In the embodiment, Z is a cyclohexyl, is not substituted or is selected from following substituting group and replaces by one or more: halogen, SO
2Rf, CONRgRh, C
1-6Alkyl, C
6-12Aralkyl, C
6-12Aryl, C
1-6Alkoxyl group, C (O) C
1-6Alkyl, C
3-10Heterocycle, hydroxyl, NRgRh, C (O) Orf or cyano group;
Rf wherein, Rg and Rh are H or C independently of one another
1-6Alkyl.
In the embodiment, Z is a piperidyl, is not substituted or is selected from following substituting group and replaces by one or more: halogen, SO
2Rf, CONRgRh, C
1-6Alkyl, C
6-12Aralkyl, C
6-12Aryl, C
1-6Alkoxyl group, C (O) C
1-6Alkyl, C (O) NHRf, C
3-10Heterocycle, hydroxyl, NRgRh, C (O) Orf or cyano group;
Rf wherein, Rg and Rh are H or C independently of one another
1-6Alkyl.
In the embodiment, Z is N (C
1-6Alkyl)-and piperidyl, be not substituted or be selected from following substituting group and replace: halogen, SO by one or more
2Rf, CONRgRh, C
1-6Alkyl, C
6-12Aralkyl, C
6-12Aryl, C
1-6Alkoxyl group, C (O) C
1-6Alkyl, C (O) NHRf, C
3-10Heterocycle, hydroxyl, NRgRh, C (O) Orf or cyano group;
Rf wherein, Rg and Rh are H or C independently of one another
1-6Alkyl.
In other embodiments: Z is a cyclohexyl, piperidyl or N (C
1-6Alkyl)-piperidyl;
Z is a cyclohexyl;
Z is a piperidyl;
Z is N (C
1-6Alkyl)-piperidyl.
In the embodiment, Z is N-methyl-piperidyl, N-ethyl-piperidyl; N-propyl group-piperidyl, N-sec.-propyl-piperidyl, N-butyl-piperidyl; N-amyl group-piperidyl, N-hexyl piperidyl, N-cyclohexyl-piperidyl; N-ethanoyl-piperidyl, N-benzyl-piperidyl, hydroxy-cyclohexyl; the oxo cyclohexyl, oxyimino cyclohexyl, aminocyclohexyl; methylsulfonyl, methylamino formyl radical or methoxyl group cyclohexyl.
In other embodiments: Z is N-methyl-piperidyl or hydroxy-cyclohexyl;
Z is N-methyl-piperidyl;
Z is N-methyl-4-piperidyl;
Z is a hydroxy-cyclohexyl;
Z is the 4-hydroxy-cyclohexyl;
Z is N-methyl-4-piperidyl.
In the embodiment, X is a 3-10 unit cycloalkyl.
In the embodiment, X is 6 yuan of cycloalkyl.
In the embodiment, X is a cyclohexyl, is not substituted or is selected from following substituting group and replaces by one or more: halogen, nitro, nitroso-group, SO
3Rf, SO
2Rf, PO
3RcRd, CONRgRh, C
1-6Alkyl, C
2-6Thiazolinyl, C
2-6Alkynyl, C
6-12Aralkyl, C
6-12Aryl, C
1-6Alkoxyl group, C
2-6Alkene oxygen base, C
2-6Alkynyloxy group, C
6-12Aryloxy, C (O) C
1-6Alkyl, C (O) C
2-6Thiazolinyl, C (O) C
2-6Alkynyl, C (O) C
6-12Aryl, C (O) C
6-12Aralkyl, C (O) NHRf, C
3-10Heterocycle, hydroxyl, NRgRh, C (O) ORf, cyano group, azido-, amidino groups or guanidine radicals;
Rf wherein, Rc, Rd, Rg and Rh are H independently of one another, C
1-6Alkyl, C
2-6Thiazolinyl, C
2-6Alkynyl, C
6-10Aryl, C
3-10Heterocycle, C
3-10Heteroaralkyl or C
6-10Aralkyl;
Or Rc and Rd form a 5-10 unit heterocycle with oxygen;
Or Rg and Rh form a 3-10 unit heterocycle with nitrogen.
In the another embodiment, X is a cyclohexyl, is not substituted or is selected from following substituting group and replaces by one or more: halogen, nitro, nitroso-group, SO
3Rf, SO
2Rf, PO
3RcRd, CONRgRh, C
1-6Alkyl, C
6-12Aralkyl, C
6-12Aryl, C
1-6Alkoxyl group, C
6-12Aryloxy, C (O) C
1-6Alkyl, C (O) C
6-12Aryl, C (O) C
6-12Aralkyl, C (O) NHRf, C
3-10Heterocycle, hydroxyl, NRgRh, C (O) ORf, cyano group, azido-, amidino groups or guanidine radicals;
Rf wherein, Rc, Rd, Rg and Rh are H independently of one another, C
1-6Alkyl, C
2-6Thiazolinyl, C
2-6Alkynyl, C
6-10Aryl, C
3-10Heterocycle, C
3-10Heteroaralkyl, C
6-10Aralkyl.
In another embodiment, X is a cyclohexyl, is not substituted or is selected from following substituting group and replaces by one or more: halogen, SO
2Rf, CONRgRh, C
1-6Alkyl, C
6-12Aralkyl, C
6-12Aryl, C
1-6Alkoxyl group, C
6-12Aryloxy, C (O) C
1-6Alkyl, C (O) C
6-12Aryl, C (O) C
6-12Aralkyl, C (O) NHRf, C
3-10Heterocycle, hydroxyl, NRgRh, C (O) ORf, cyano group or azido-;
Rf wherein, Rc, Rd, Rg and Rh are H independently of one another, C
1-6Alkyl, C
6-10Aryl, C
3-10Heterocycle, C
3-10Heteroaralkyl or C
6-10Aralkyl.
In the embodiment, X is a cyclohexyl, by one or more following substituting group replacement: C that are selected from
1-6Alkyl, halogen, C
2-6Thiazolinyl, C
2-6Alkynyl or C
1-6Alkoxyl group.
In other embodiments: X is by C
1-6The cyclohexyl that alkyl replaces;
The cyclohexyl that X is replaced by the C1-3 alkyl;
X is 4-methyl-cyclohexyl base or 2-hydroxy-4-methyl-cyclohexyl;
X is the 4-methylcyclohexyl.
In the embodiment, Y is a 6-10 unit aryl.
In the embodiment, Y is a phenyl, is not substituted or is selected from following substituting group and replaces by one or more: halogen, nitro, nitroso-group, SO
3Rf, SO
2Rf, PO
3RcRd, CONRgRh, C
1-6Alkyl, C
2-6Thiazolinyl, C
2-6Alkynyl, C
6-12Aralkyl, C
6-12Aryl, C
1-6Alkoxyl group, C
2-6Alkene oxygen base, C
2-6Alkynyloxy group, C
6-12Aryloxy, C (O) C
1-6Alkyl, C (O) C
2-6Thiazolinyl, C (O) C
2-6Alkynyl, C (O) C
6-12Aryl, C (O) C
6-12Aralkyl, C (O) NHRf, C
3-10Heterocycle, hydroxyl, NRgRh, C (O) ORf, cyano group, azido-, amidino groups or guanidine radicals;
Rf wherein, Rc, Rd, Rg and Rh are H independently of one another, C
1-6Alkyl, C
2-6Thiazolinyl, C
2-6Alkynyl, C
6-10Aryl, C
3-10Heterocycle, C
3-10Heteroaralkyl or C
6-10Aralkyl;
Or Rc and Rd form a 5-10 unit heterocycle with oxygen;
Or Rg and Rh and nitrogen form a 3-10 unit heterocycle together.
In the another embodiment, Y is a phenyl, is not substituted or is selected from following substituting group and replaces by one or more: halogen, nitro, nitroso-group, SO
3Rf, SO
2Rf, PO3RcRd, CONRgRh, C
1-6Alkyl, C
2-6Thiazolinyl, C
2-6Alkynyl, C
6-12Aralkyl, C
6-12Aryl, C
1-6Alkoxyl group, C
2-6Alkene oxygen base, C
2-6Alkynyloxy group, C
6-12Aryloxy, C (O) C
1-6Alkyl, C (O) C
2-6Thiazolinyl, C (O) C
2-6Alkynyl, C (O) C
6-12Aryl, C (O) C
6-12Aralkyl, C (O) NHRf, C
3-10Heterocycle, hydroxyl, NRgRh, C (O) ORf, cyano group, azido-, amidino groups or guanidine radicals;
Rf wherein, Rc, Rd, Rg and Rh are H independently of one another, C
1-6Alkyl, C
2-6Thiazolinyl, C
2-6Alkynyl, C
6-10Aryl, C
3-10Heterocycle, C
3-10Heteroaralkyl or C
6-10Aralkyl.
In another embodiment, Y is a phenyl, is not substituted or is selected from following substituting group and replaces by one or more: halogen, nitro, nitroso-group, SO
3Rf, SO
2Rf, PO
3RcRd, CONRgRh, C
1-6Alkyl, C
6-12Aralkyl, C
6-12Aryl, C
1-6Alkoxyl group, C
6-12Aryloxy, C (O) C
1-6Alkyl, C (O) C
6-12Aryl, C (O) C
6-12Aralkyl, C (O) NHRf, C
3-10Heterocycle, hydroxyl, NRgRh, C (O) ORf, cyano group, azido-, amidino groups or guanidine radicals;
Rf wherein, Rc, Rd, Rg and Rh are H independently of one another,
1-6Alkyl, C
2-6Thiazolinyl, C
2-6Alkynyl, C
6-10Aryl,
3-10Heterocycle, C
3-10Heteroaralkyl or C
6-10Aralkyl.
In the embodiment, Y is a phenyl, is not substituted or is selected from following substituting group and replaces by one or more: halogen, nitro, SO
2Rf, CONRgRh, C
1-6Alkyl, C
6-12Aralkyl, C
6-12Aryl, C
1-6Alkoxyl group, C
6-12Aryloxy, C (O) C
1-6Alkyl, C (O) C
6-12Aryl, C (O) C
6-12Aralkyl, C (O) NHRf, C
3-10Heterocycle, hydroxyl, NRgRh, C (O) ORf, cyano group, amidino groups or guanidine radicals;
Rf wherein, Rg and Rh are H independently of one another, C
1-6Alkyl, C
6-10Aryl, C
3-10Heterocycle, C
3-10Heteroaralkyl or C
6-10Aralkyl.
In the embodiment, Y is a phenyl, is selected from following substituting group and replaces by one or more: halogen, nitro, SO
2Rf, C
1-6Alkyl, C
1-6Alkoxyl group, C (O) C
1-6Alkyl, C (O) ORf, cyano group or azido-.
In the embodiment, Y is a phenyl, is selected from following substituting group and replaces by one or more: halogen, nitro, C
1-6Alkyl, C
1-6Alkoxyl group or cyano group.
In other embodiments: the phenyl that Y is replaced by one or more halogens;
Y is by one or more C
1-6The phenyl that alkoxyl group replaces;
The phenyl that Y is replaced by one or more methoxyl groups;
Y is by one or more C
1-6The phenyl that alkyl replaces;
Y is by one or more methyl substituted phenyl;
Y is a phenyl;
Y is the 3-fluorophenyl, 4-fluorophenyl 4-chloro-phenyl-, 4-cyano-phenyl, 4-p-methoxy-phenyl, 4-nitrophenyl or p-methylphenyl.
In the embodiment, P is the 1-3 integer, and q is the 0-2 integer;
In other embodiments: P is 2, and q is 2;
P is 3, and q is 2;
P is 1, and q is 2;
P is 1, and q is 1;
P is 2, and q is 1;
P is 3, and q is 1;
P is 1, and q is 0;
P is 2, and q is 0;
P is 3, and q is 0.
In the embodiment, W is CR
10R
11S (O) n, O or NR
12
N wherein, R
10R
11And R
12By definition herein.
In other embodiments: W is CR
10R
11Or NR
12
R wherein
10R
11And R
12By definition herein;
W is CR
10R
11
R wherein
10And R
11By definition herein;
W is NR
12
R wherein
12By definition herein;
W is 0;
W is S (O) n;
Wherein n is by definition herein.
In the embodiment, R
10And R
11Be independently selected from: H, C
1-6Alkyl, C
6-0Aryl, C
3-10Heterocycle, C
3-10Heteroaralkyl, C
6-10Aralkyl, C (O)-C
1-6Alkyl, C
1-6Alkoxyl group, hydroxyl or formyl radical;
Or R
10And R
11Together formation=0.=S or=N-Ra, wherein Ra is H, hydroxyl or C
1-6Alkyl.
In the embodiment, R
10Be H, C
1-6Alkyl, C
6-10Aryl, C
3-10Heterocycle, C
3-10Heteroaralkyl, C
6-10Aralkyl, C (O)-C
1-6Alkyl, C
1-6Alkoxyl group, hydroxyl or formyl radical; And R
11Be H.
In the embodiment, R
10Be C
1-6Alkyl, C
6-10Aralkyl, C (O)-C
1-6Alkyl, C
1-6Alkoxyl group, hydroxyl or formyl radical; And R
11Be H.
In the embodiment, R
10Be C
1-3Alkyl, C
6-10Aralkyl, C (O)-C
1-3Alkyl, C
1-3Alkoxyl group, hydroxyl or formyl radical; And R
11Be H.
In the embodiment, R
10Be selected from: methyl, ethyl, propyl group, isopropyl benzyl, ethanoyl, hydroxyl or formyl radical; And R
11Be H.
In the embodiment, R
10And R
11Form=0 together ,=S or=N-Ra, wherein Ra is H, hydroxyl or C
1-6Alkyl.
In the embodiment, R
10And R
11Form=0 together.
In the another embodiment, R
10And R
11Formation=S together.
In the another embodiment, R
10And R
11Formation=N-Ra together, wherein Ra is H, hydroxyl or C
1-6Alkyl.
In the embodiment, Ra is selected from H, hydroxyl, methyl, ethyl, propyl group or sec.-propyl.
In the embodiment, R
13Be one or more optional substituting groups, be selected from halogen independently of one another, nitro, nitroso-group, SO
3Rf, SO
2Rf, PO
3RcRd, CONRgRh, C
1-6Alkyl, C
2-6Thiazolinyl, C
2-6Alkynyl, C
2-12Aralkyl, C
6-12Aryl, C
1-6Alkoxyl group, C
2-6Alkene oxygen base, C
2-6Alkynyloxy group, C
6-12Aryloxy, C (O) C
1-6Alkyl, C (O) C
2-6Thiazolinyl, C (O) C
2-6Alkynyl, C (O) C
6-12Aryl, C (O) C
6-12Aralkyl, C
3-10Heterocycle, hydroxyl, NRgRh, C (O) ORf, cyano group, azido-, amidino groups or guanidine radicals;
Rf wherein, Rc, Rd, Rg and Rh are H independently of one another, C
1-6Alkyl, C
2-6Thiazolinyl, Cs-e alkynyl, C
6-10Aryl, C
3-10Heterocycle, C
3-10Heteroaralkyl or C
6-10Aralkyl;
Or Rc and Rd form a 5-10 unit heterocycle with oxygen;
Or Rg and Rh form one with nitrogen
3-10Unit's heterocycle.
In the embodiment, R
13Be one or more optional substituting groups, be selected from independently of one another: halogen, nitro, SO
2Rf, CONRgRh, C
1-6Alkyl, C
6-12Aralkyl, C
6-12Aryl, C
1-6Alkoxyl group, C (O) C
1-6Alkyl, C
3-10Heterocycle, hydroxyl, NRgRh, C (O) ORf, cyano group or azido-;
Rf wherein, Rg and Rh are by definition herein.
In the embodiment, R
13Be one or more optional substituting groups, they are independently selected from: halogen, nitro, SO
2Rf, CONRgRh, C
1-3Alkyl, C
6-12Aralkyl, C
6Aryl, C
1-3Alkoxyl group, C (O) C
1-3Alkyl, C3-6 heterocycle, hydroxyl, NRgRh, C (O) ORf, cyano group or azido-;
Rf wherein, Rg and Rh press this paper definition.
In the embodiment, R
13Be one or more optional substituting groups, be selected from independently of one another: halogen, nitro, SO
2CH
3, CONH
2, CONHCH
3, CONH (CH
3)
2, methyl, ethyl, propyl group, sec.-propyl, benzyl, phenyl, ethanoyl, methoxyl group, oxyethyl group, propoxy-, isopropoxy, piperidyl, piperazinyl, pyrrolidyl, azetidinyl, aziridinyl, pyridyl ， alkyl dioxin, dioxolanyl, azepanyl, hydroxyl, NH
2, N (H) CH
3, NH (CH
3)
2, cyano group or azido-;
Rf wherein, Rg and Rh are by definition herein.
It will be apparent to those skilled in the art that R
13Can be connected to any position of ring.Those skilled in the art also understand R
13The valence mumber of annular atoms is depended in position on described ring, and observes the conventional chemical rule.
In the embodiment, the invention provides the compound of following structural formula representative, or its pharmacy acceptable salt;
In the formula;
Z is a cyclohexyl, is not substituted or independently is selected from following substituting group and replaces by one or more: halogen, SO
2Rf, CONRgRh, C
1-6Alkyl, C
6-12Aralkyl, C
6-12Aryl, C
1-6Alkoxyl group, C (O) C
1-6Alkyl, C
3-10Heterocycle, hydroxyl, NRgRh, C (O) ORf or cyano group;
Wherein, Rf, Rg and Rh are H or C independently of one another
1-6Alkyl;
Y is a phenyl, is not substituted or independently is selected from following substituting group and replaces by one or more: halogen, nitro, SO2Rf, CONRgRh, C
1-6Alkyl, C
6-12Aralkyl, C
6-12Aryl, C
1-6Alkoxyl group, C
6-12Aryloxy, C (O) C
1-6Alkyl, C (O) C
6-12Aryl, C (O) C
6-12Aralkyl, C (O) NHRf, C
3-10Heterocycle, hydroxyl, NRgRh, C (O) ORf, cyano group, amidino groups or guanidine radicals;
Wherein, Rf, Rg and Rh are H independently of one another, C
1-6Alkyl, C
6-10Aryl, C
3-10Heterocycle, C
3-10Heteroaralkyl or C
6-10Aralkyl;
X is a cyclohexyl, is not substituted or independently is selected from following substituting group and replaces by one or more: halogen, SOZRf, CONRgRh, C
1-6Alkyl, C
6-12Aralkyl, C
6-12Aryl, C
1-6Alkoxyl group, C
6-12Aryloxy, C (O) C
1-6Alkyl, C (O) C
6-12Aryl, C (O) C aralkyl, C (O) NHRf, C
3-10Heterocycle, hydroxyl, NRgRh, C (O) ORf, cyano group or azido-;
Wherein, Rf, Rc, Rd, Rg and Rh are H independently of one another, C
1-6Alkyl, C
6-10Aryl, C
3-10Heterocycle, C
3-10Heteroaralkyl or C
6-10Aralkyl;
M is 0;
Condition is when Y is unsubstituted phenyl, and then X is not the 4-methylcyclohexane.
On the one hand, compounds provided by the invention comprises:
Compound 1:3-{[(2-carboxyl-5-phenyl-benzene sulphur-3-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-methyl }-piperidines; Trifluoroacetate;
Compound 2:2-{[(2-carboxyl-5-phenyl-benzene sulphur-3-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-methyl }-piperidines; Trifluoroacetate;
Compound 3:3-[(4-methyl-cyclohexyl alkyl carbonyl)-pyridin-3-yl methyl-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 4:3-[(4-methyl-cyclohexyl alkyl carbonyl)-pyridin-4-yl methyl-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 5:5-(3-fluoro-phenyl)-3-[sec.-propyl-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-thiophene-2-carboxylic acid;
Compound 6:3-[AZEPAN-4-base-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 7:3-[(2,4-two chloro-benzoyls)-[1,3] dioxolane-2-ylmethyl-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 8:3-[[1,3] dioxolane-2-ylmethyl-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 9:3-[(1-fluoro-4-methyl-cyclohexyl alkyl carbonyl)-sec.-propyl-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 10:3-[(1-fluoro-4-methyl-cyclohexyl alkyl carbonyl)-sec.-propyl-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 11: chlorination 4-[(2-carboxyl-5-phenyl-benzene sulphur-3-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-1-methyl-piperidines;
Compound 12:3-[(2-acetylamino-4-methyl-cyclohexyl alkyl carbonyl)-sec.-propyl-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 13:3-[(4-methyl-cyclohexyl alkyl carbonyl)-(4-oxygen-cyclohexyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 14:3-[(4-methyl-cyclohexyl alkyl carbonyl)-pyridine-2-ylmethyl-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 15:3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 16:3-[(4-oxyimino-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 17:3-[sec.-propyl-(4-methyl-cyclohexyl-3-alkene carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 18:3-[(1-azido methyl-2-methyl-butyl)-(2,4-two chloro-benzoyls)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 19: trifluoroacetic acid 2-[(2 carboxyl-5-phenyl-benzene sulphur-3-yl)-(2-chloro-benzoyl)-amino]-3-methyl-amyl group-ammonium;
Compound 20:3-[(1-amino methyl-2-methyl-butyl)-(2,4-two chloro-benzoyls)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 21:{2-[(2-carboxyl-5-phenyl-benzene sulphur-3-yl)-(2,4-two chloro-benzoyls)-amino]-propyl group }-trimethylammonium-ammonium; Trifluoroacetate;
Compound 22:3-[sec.-propyl-(the 5-methyl-[1,3] diox-2-carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 23: chlorination 4-[[2-carboxyl-5-(4-fluoro-phenyl)-benzene sulphur-3-yl]-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-1-methyl-piperidines;
Compound 24:5-(4-fluoro-phenyl)-3-[(2-hydroxy-4-methyl-hexanaphthene carbonyl)-sec.-propyl-amino]-thiophene-2-carboxylic acid;
Compound 25:3-[(4-methoxyimino-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 26:5-(4-fluoro-phenyl)-3-[sec.-propyl-(4-methyl-cyclohexyl-1-alkene carbonyl)-amino]-thiophene-2-carboxylic acid;
Compound 27:3-[sec.-propyl-(5-methyl-tetrahydrochysene-pyrans-2-carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 28:3-[sec.-propyl-(4-methylene radical-hexanaphthene carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 29:3-[sec.-propyl-(5-methyl-tetrahydrochysene-pyrans-2-carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 30:3-[sec.-propyl-(5-methyl-3.6-dihydro-2H-pyrans-2-carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 31:3-[(2-hydroxy-4-methyl-hexanaphthene carbonyl)-(tetrahydrochysene-pyrans-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 32:3-[(2-azido--1-methyl-ethyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 33:3-[(4-methyl-cyclohexyl alkyl carbonyl)-(1-methyl-piperidin-4-yl methyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 34:3-[(4-methyl-cyclohexyl alkyl carbonyl)-(tetrahydrochysene-thiapyran-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 35: chlorination 3-{[(2-carboxyl-5-phenyl-benzene sulphur-3-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-methyl }-1-methyl-piperidines;
Compound 36:3-[(2-amino-1-methyl-ethyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 37:3-[(4-methyl-cyclohexyl alkyl carbonyl)-(1,1-dioxy-tetrahydrochysene-thiapyran-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 38: chlorination 4-{[(2-carboxyl-5-phenyl-benzene sulphur-3-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-methyl }-1-methyl-piperidines;
Compound 39:3-[(1-ethyl-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 40:3-[(1-sec.-propyl-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 41:3-[(4-methyl-cyclohexyl alkyl carbonyl)-piperidin-4-yl-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 42:3-[[1-(4-methoxyl group-benzyl)-2-oxygen-piperidin-4-yl]-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 43:3-[(2-azido--1-methyl-ethyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 44:5-(3-fluoro-phenyl)-3-[(2-hydroxy-4-methyl-hexanaphthene carbonyl)-sec.-propyl-amino]-thiophene-2-carboxylic acid;
Compound 45: chlorination 4-[(2-carboxyl-5-tolyl-benzene sulphur-3-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-1-methyl-piperidines;
Compound 46:3-[(4-methoxyl group-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 47:3-[(4-methyl-cyclohexyl alkyl carbonyl)-(4-hydroxyl-cyclohexyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 48:3-[(1-ethanoyl-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 49: chlorination 4-[(2-carboxyl-5-phenyl-benzene sulphur-3-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-1-methyl-AZEPANIUM;
Compound 50:5-(4-fluoro-phenyl)-3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-thiophene-2-carboxylic acid;
Compound 51:5-(3-fluoro-phenyl)-3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-thiophene-2-carboxylic acid;
Compound 52:3-[(1-benzyl-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 53:5-(4-fluoro-phenyl)-3-[sec.-propyl-(4-methyl-cyclohexyl-3-alkene carbonyl)-amino]-thiophene-2-carboxylic acid;
Compound 54:4-[[2-carboxyl-5-(3-fluoro-phenyl)-benzene sulphur-3-yl]-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-1-methyl-piperidines; Muriate
Compound 55:4-[[2-carboxyl-5-(4-methoxyl group-phenyl)-benzene sulphur-3-yl]-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-1-methyl-piperidines; Muriate;
Compound 56:4-[[2-carboxyl-5-(4-nitro-phenyl)-benzene sulphur-3-yl]-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-1-methyl-piperidines; Muriate;
Compound 57: chlorination 4-[[2-carboxyl-5-(4-chloro-phenyl)-benzene sulphur-3-yl]-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-1-methyl-piperidines;
Compound 58: chlorination 4-[[2-carboxyl-5-(4-cyano group-phenyl)-benzene sulphur-3-yl]-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-1-methyl-piperidines;
Compound 59:5-(4-chloro-phenyl)-3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-thiophene-2-carboxylic acid;
Compound 60:3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-(4-methoxyl group-phenyl)-thiophene-2-carboxylic acid;
Compound 61:5-(4-cyano group-phenyl)-3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-thiophene-2-carboxylic acid;
Compound 62:3-[(2-hydroxy-4-methyl-hexanaphthene carbonyl)-sec.-propyl-amino]-5-(4-methoxyl group-phenyl)-thiophene-2-carboxylic acid;
Compound 63:3-[(1-formyl radical-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 64:3-[N ', N '-dimethyl-N-(4-methyl-cyclohexyl alkyl carbonyl)-diazanyl]-5-phenyl-thiophene-2-carboxylic acid;
Compound 65:3-[(4-methyl-cyclohexyl alkyl carbonyl)-(1-methyl isophthalic acid-oxygen-piperidin-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 66:3-[(4-methyl-cyclohexyl alkyl carbonyl)-(1-methyl isophthalic acid-oxygen-piperidin-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 67:3-[(2-amino-cyclohexyl)-(2,4-two chloro-benzoyls)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 68:3-[(4-methyl-cyclohexyl alkyl carbonyl)-(1-oxygen-tetrahydrochysene-thiapyran-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 69:5-(4-fluorophenyl)-((4-methyl-cyclohexyl alkyl carbonyl)-1-(methyl-piperidines-3-ylmethyl)-amino)-thiophene-2-carboxylic acid;
Compound 70:3-[(1-methylsulfonyl-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 71:3-[(1-methylamino formyl radical-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 72:3-[N-(2,4-two chloro-benzoyls)-N ', N '-dimethyl-diazanyl]-5-phenyl-thiophene-2-carboxylic acid;
Or its pharmacy acceptable salt.
On the one hand, compounds provided by the invention comprises:
Compound 73:5-(4-fluoro-phenyl)-3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-thiophene-2-carboxylic acid;
Compound 74:3-[(1-methylamino formyl radical-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 75:3-[(4-methyl-cyclohexyl alkyl carbonyl)-(1-methyl-2-oxygen-piperidin-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 76:3-[(4-carboxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 77:3-[(1-cyano group-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 78:3-[(4-carboxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 79:5-(3.4-two fluoro-phenyl)-3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-thiophene-2-carboxylic acid;
Compound 80:5 '-ethanoyl-4-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-[2.2 '] hexichol sulfenyl-5-carboxylic acid;
Compound 81:3-[(1-formamyl-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 82:3-[(4-methyl-cyclohexyl alkyl carbonyl)-(7-oxygen-AZEPAN-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 83:3-[(1-oxamoyl base-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 84:3-[ethyl-(4-methyl-benzoyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 85:5-(4-ethanoyl-phenyl)-3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-thiophene-2-carboxylic acid;
Compound 86:3-[(4-hydroxy-4-methyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 87:3-[(3-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 88:3-[(4-hydroxy-4-methyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 89:3-[(3-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 90:3-[(3-hydroxyl-cyclopentyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Or its pharmacy acceptable salt.
In the embodiment, novel 3-[(6-provided by the invention unit cycloalkyl-carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid compound is selected from following compounds:
Compound 1:3-{[(2-carboxyl-5-phenyl-benzene sulphur-3-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-methyl }-piperidines; Trifluoroacetate;
Compound 2:2-([(2-carboxyl-5-phenyl-benzene sulphur-3-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-methyl)-piperidines; Trifluoroacetate;
Compound 3:3-[(4-methyl-cyclohexyl alkyl carbonyl)-pyridin-3-yl methyl-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 4:3-[(4-methyl-cyclohexyl alkyl carbonyl)-pyridin-4-yl methyl-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 6:3-[AZEPAN-4-base-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 8:3-[[1,3] dioxolane-2-ylmethyl-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 9:3-[(1-fluoro-4-methyl-cyclohexyl alkyl carbonyl)-sec.-propyl-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 10:3-[(1-fluoro-4-methyl-cyclohexyl alkyl carbonyl)-sec.-propyl-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 11: chlorination 4-[(2-carboxyl-5-phenyl-benzene sulphur-3-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-1-methyl-piperidines;
Compound 12:3-[(2-acetylamino-4-methyl-cyclohexyl alkyl carbonyl)-sec.-propyl-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 13:3-[(4-methyl-cyclohexyl alkyl carbonyl)-(4-oxygen-cyclohexyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 14:3-[(4-methyl-cyclohexyl alkyl carbonyl)-pyridine-2-ylmethyl-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 15:3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 16:3-[(4-oxyimino-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 17:3-[sec.-propyl-(4-methyl-cyclohexyl-3-alkene carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 25:3-[(4-methoxyimino-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 28:3-[sec.-propyl-(4-methylene radical-hexanaphthene carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 31:3-[(2-hydroxy-4-methyl-hexanaphthene carbonyl)-(tetrahydrochysene-pyrans-4-yl)-amino I-5-phenyl-thiophene-2-carboxylic acid;
Compound 32:3-[(2-azido--1-methyl-ethyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 33:3-[(4-methyl-cyclohexyl alkyl carbonyl)-(1-methyl-piperidin-4-yl methyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 34:3-[(4-methyl-cyclohexyl alkyl carbonyl)-(tetrahydrochysene-thiapyran-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 35: chlorination 3-{[(2-carboxyl-5-phenyl-benzene sulphur-3-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-methyl }-1-methyl--piperidines;
Compound 36:3-[(2-amino-1-methyl-ethyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 37:3-[(4-methyl-cyclohexyl alkyl carbonyl)-(1-oxygen-six hydrogen-thiapyran-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 38: chlorination 4-{[(2-carboxyl-5-phenyl-benzene sulphur-3-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-methyl }-1-methyl-piperidines;
Compound 39:3-[(1-ethyl-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 40:3-[(1-sec.-propyl-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 41:3-[(4-methyl-cyclohexyl alkyl carbonyl)-piperidin-4-yl-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 42:3-[[1-(4-methoxyl group-benzyl)-2-oxygen-piperidin-4-yl]-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 43:3-[(2-azido--1-methyl-ethyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 46:3-[(4-methoxyl group-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 47:3-[(4-methyl-cyclohexyl alkyl carbonyl)-(4-methyl-cyclohexyl base)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 48:3-[(1-ethanoyl-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 49: chlorination 4-[(2-carboxyl-5-phenyl-benzene sulphur-3-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-1-methyl-AZEPANIUM;
Compound 52:3-[(1-benzyl-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 63:3-[(1-formyl radical-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 64:3-[N ', N '-dimethyl-N-(4-methyl-cyclohexyl alkyl carbonyl)-diazanyl]-5-phenyl-thiophene-2-carboxylic acid;
Compound 65:3-[(4-methyl-cyclohexyl alkyl carbonyl)-(1-methyl isophthalic acid-oxygen-piperidin-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 66:3-[(4-methyl-cyclohexyl alkyl carbonyl)-(1-methyl isophthalic acid-oxygen-piperidin-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 68:3-[(4-methyl-cyclohexyl alkyl carbonyl)-(1-oxygen-six hydrogen-thiapyran-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 70:3-[(1-methylsulfonyl-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 71:3-[(1-methylamino formyl radical-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 74:3-[(1-methylamino formyl radical-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 75:3-[(4-methyl-cyclohexyl alkyl carbonyl)-(1-methyl-2-oxygen-piperidin-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 76:3-[(4-carboxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 77:3-[(1-cyano group-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 78:3-[(4-carboxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 81:3-[(1-formamyl-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 82:3-[(4-methyl-cyclohexyl alkyl carbonyl)-(7-oxygen-AZEPAN-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 83:3-[(1-oxamoyl base-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 86:3-[(4-hydroxy-4-methyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 87:3-[(3-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 88; 3-[(4-hydroxy-4-methyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 89:3-[(3-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 90:3-[(3-hydroxyl-cyclopentyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Or its pharmacy acceptable salt.
In the embodiment, novel 3-[(4-methyl-cyclohexyl alkane-carbonyl provided by the invention)-amino]-5-phenyl-thiophene-2-carboxylic acid compound.
In the embodiment, novelty provided by the invention have 3-[(be not substituted or replace-benzoyl)-amino]-5-phenyl-thiophene-2-carboxylic acid compound is selected from following compounds:
Compound 7:3-[(2,4-two chloro-benzoyls)-[1,3] dioxolane-2-ylmethyl-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 18:3-[(1-azido methyl-2-methyl-butyl)-(2,4-two chloro-benzoyls)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 19:2-[(2-carboxyl-5-phenyl-benzene sulphur-3-yl)-(2-chloro-benzoyl)-amino]-3-methyl-amyl group-ammonium, trifluoroacetate;
Compound 20:3-[(1-amino methyl-2-methyl-butyl)-(2,4-two chloro-benzoyls)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 21:{2-[(2-carboxyl-5-phenyl-benzene sulphur-3-yl)-(2,4-two chloro-benzoyls)-amino]-propyl group }-trimethylammonium-ammonium, trifluoroacetate;
Compound 67:3-[(2-amino-cyclohexyl)-(2,4-two chloro-benzoyls)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 72:3-[N-(2,4-two chloro-benzoyls)-N ', N '-dimethyl-diazanyl]-5-phenyl-thiophene-2-carboxylic acid;
Compound 84:3-[ethyl-(4-methyl-benzoyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Or its pharmacy acceptable salt.
In the embodiment, novel 3-[(6-provided by the invention unit heterocycle-2-carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid compound is selected from following compounds:
Compound 22:3-[sec.-propyl-(the 5-methyl-[1,3] diox-2-carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 27:3-[sec.-propyl-(5-methyl-tetrahydrochysene-pyrans-2-carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 29:3-[sec.-propyl-(5-methyl-tetrahydrochysene-pyrans-2-carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Compound 30:3-[sec.-propyl-(5-methyl-3.6-dihydro-2H-pyrans-2-carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
Or its pharmacy acceptable salt.
In the embodiment, virus infection is selected from flaviviridae infections.
In the embodiment, flaviviridae infections is selected from hepatitis C virus (HCV), bovine viral diarrhea virus (BVDV), Pestivirus suis, dengue fever virus, japanese encephalitis virus and yellow fever virus.
In another embodiment, flaviviridae infections is an infection with hepatitis C virus.
In the embodiment, the invention provides the method that a kind of treatment or prevention host flavivirus infect, comprise giving the The compounds of this invention at least a as herein described that the host treats significant quantity.
In the embodiment; the invention provides the method that a kind of treatment or prevention host flavivirus infect; comprise and give the The compounds of this invention at least a as herein described that the host treats significant quantity; also comprise and give at least a other the following preparations that are selected from: virus serine protease class inhibitor; viral polymerase inhibitors; virus helicase inhibitor; immunomodulator; antioxidant; antiseptic-germicide; therapeutic vaccine, hepatoprotective (hepatoprotectant agent) or antisense agents.
In the embodiment, described other preparations are interferon alphas, virazole, Silymarin, il-1 2. amantadines, rnase, thymosin, N-acetylcysteine or ciclosporin.
In the embodiment, it is infection with hepatitis C virus (HCV) that yellow heat belongs to virus infection.
In the embodiment, the invention provides a kind of pharmaceutical composition, it comprises at least a The compounds of this invention as herein described and at least a pharmaceutically acceptable carrier or vehicle.
In the embodiment; the invention provides a kind of pharmaceutical composition; it comprises at least a The compounds of this invention as herein described and at least a pharmaceutically acceptable carrier or vehicle, also comprises at least a other the following preparations that are selected from: virus serine protease class inhibitor, viral polymerase inhibitors; virus helicase inhibitor; immunomodulator, antioxidant, antiseptic-germicide; therapeutic vaccine, hepatoprotective or antisense agents.
In another embodiment, described other preparations are interferon alphas, virazole, Silymarin, recombinant interleukin-1 2, amantadine, rnase, thymosin, N-acetylcysteine or ciclosporin.
In the embodiment, virus serine protease class inhibitor is a Flavivirus serine stretch protein enzyme inhibitor.
In the embodiment, viral polymerase inhibitors is the Flavivirus AG14361.
In the embodiment, viral helicase inhibitor is a Flavivirus helicase inhibitor.
In other embodiments: virus serine protease class inhibitor is a HCV serine stretch protein enzyme inhibitor; Viral polymerase inhibitors is the HCV AG14361; Virus helicase inhibitor is a HCV helicase inhibitor.
In the embodiment, a kind of inhibition is provided or reduces the active method of varial polymerases among the host, comprise the The compounds of this invention as herein described for the treatment of significant quantity.
In the embodiment, a kind of inhibition is provided or reduces the active method of varial polymerases among the host, comprise the The compounds of this invention as herein described for the treatment of significant quantity, also comprise giving one or more viral polymerase inhibitors.
In the embodiment, varial polymerases is the flavivirus polysaccharase.
In the embodiment, varial polymerases is the RNA-polysaccharase that RNA-relies on.
In the embodiment, varial polymerases is the HCV polysaccharase.
In the embodiment; a kind of Combined Preparation is provided; comprise at least a The compounds of this invention as herein described and be selected from other following preparations: virus serine protease class inhibitor with one or more; viral polymerase inhibitors and viral helicase inhibitor; immunomodulator, antioxidant, antiseptic-germicide; therapeutic vaccine, hepatoprotective or antisense agents.
In the embodiment, give described compound and other preparations in succession.
In the embodiment, give described compound and other preparations simultaneously.
Above-mentioned Combined Preparation usually can pharmaceutical preparation form use, therefore comprising as mentioned above, the pharmaceutical preparation of coupling medicine and pharmaceutically acceptable carrier is another aspect of the present invention.
Each component of using in the inventive method or the combination can give or give simultaneously by the separated drug preparation in succession, or gives with combination preparation.
In the embodiment, the invention provides the application of The compounds of this invention described herein in treatment or the infection of prevention host flavivirus.
In the embodiment, the invention provides The compounds of this invention as herein described in manufacturing treatment or the yellow hot application that belongs in the medicine that infects of prevention host virus.
In the embodiment, the invention provides The compounds of this invention as herein described application in the varial polymerases activity in inhibition or reduction host.
Those skilled in the art can understand, and compound of the present invention can contain chiral centre.Therefore described structural formula compound can two kinds of different optical isomer forms (i.e. (+) or (-) enantiomorph) exist.Enantiomorph that all are such and composition thereof comprises that racemic mixture is included within the scope of the invention.The method that single optical isomer or enantiomorph can all be known by this area, chirality HPLC for example, enzyme is differentiated and the auxiliary acquisition of chirality.
Preferably, The compounds of this invention is with at least 95%, and better at least 97%, best a kind of enantiomorph of at least 99%, and do not have the form of its corresponding enantiomorph to provide.
Better, The compounds of this invention is (+) enantiomorph of at least 95%, and does not have the form of corresponding (-) enantiomorph.
Better, The compounds of this invention is (+) enantiomorph of at least 97%, and does not have the form of corresponding (-) enantiomorph.
Better, The compounds of this invention is (+) enantiomorph of at least 99%, and does not have the form of corresponding (-) enantiomorph.
In the better implement mode, The compounds of this invention is at least 95% (-) enantiomorph, and does not have the form of corresponding (+) enantiomorph.Best, The compounds of this invention is at least 97% (-) enantiomorph, not corresponding (+) enantiomeric form.
Better, The compounds of this invention is at least 99% (-) enantiomorph, and not corresponding (+) enantiomeric form.
Should also be understood that The compounds of this invention can contain more than one chiral centre.Therefore the diastereomer form that the compound of described structural formula can be different exists.All such diastereomers and its mixture are included within the scope of the invention.The method that single diastereomer can all be known by this area, as HPLC, crystallization and chromatography obtain.
The pharmacy acceptable salt of The compounds of this invention also is provided.The pharmacy acceptable salt of term compound refers to produce from pharmaceutically acceptable inorganic and organic acid and inorganic and those salt organic bases.The example of appropriate acid comprises hydrochloric acid, Hydrogen bromide, sulfuric acid, nitric acid, perchloric acid, fumaric acid, toxilic acid, phosphoric acid, oxyacetic acid, lactic acid, Whitfield's ointment, succsinic acid, toluene-right-sulfonic acid, tartrate, acetate, trifluoroacetic acid, citric acid, methylsulfonic acid, formic acid, phenylformic acid, propanedioic acid, naphthalene-2-sulfonic acid and Phenylsulfonic acid.Other acid although itself be not pharmaceutically acceptable, can be used as intermediate as oxalic acid in obtaining The compounds of this invention and its pharmaceutically-acceptable acid addition.
Generation comprises basic metal (as sodium, lithium, potassium) from the salt of appropriate base, alkaline-earth metal (as magnesium), and (wherein R is C for ammonium and NR4+
1-4Alkyl) salt.
The object of reference of following The compounds of this invention comprises compound and its pharmacy acceptable salt.
In one embodiment of the present invention, pharmacy acceptable salt is a sodium salt.
In one embodiment of the present invention, pharmacy acceptable salt is a lithium salts.
In one embodiment of the present invention, pharmacy acceptable salt is a sylvite.
The applicant also submits on June 11st, 2002 and waits to examine US regular application 10/166,031. title and be: " compound and the using method of treatment or prevention of flavivirus infections ", it is for referencial use to fit into this paper in it.
Unless otherwise defined, all technical terms used herein and scientific terminology have the identical meanings that those of ordinary skills understand.All publications that this paper addresses, patent application, patent and other reference are all included this paper reference in.
In conflict situations, this specification sheets comprises that definition will play dominating role (control).In addition, material, method and embodiment be explanation just, is not construed as limiting.
Straight or branched hydrocarbon part represented in the term that uses among the application " alkyl ", can choose wantonly by one or more to be selected from following group and to replace: halogen, nitro, nitroso-group, SO
3R
12PO
3RcRd, CONR
13R
14C
1-6Alkyl, C
2-6Thiazolinyl, C
2-6Alkynyl, C
6-12Aralkyl, C
6-12Aryl, C
1-6Alkoxyl group, C
2-6Alkene oxygen base, C
2-6Alkynyloxy group, C
6-12Aryloxy, C (O) C
1-6Alkyl, C (O) C
2-6Thiazolinyl, C (O) C
2-6Alkynyl, C (O) C
6-12Aryl, C (O) C
6-12Aralkyl, C
3-10Heterocycle, hydroxyl, NR
13R
14C (O) OR
12Cyano group, azido-, amidino groups or guanidine radicals;
R wherein
12Rc, Rd, R
13And R
14Be selected from independently of one another: H, C
1-12Alkyl, C
2-12Thiazolinyl, C
2-12Alkynyl, C
6-14Aryl, C
3-12Heterocycle, C
3-18Heteroaralkyl, C
6-18Aralkyl;
Or Rc and Rd form a 5-10 unit heterocycle with oxygen;
Or R
13And R
14Form a 3-10 unit heterocycle with nitrogen.
The examples of alkyl groups that is suitable for comprises: sec.-propyl, ethyl, fluorine hexyl or cyclopropyl.The term alkyl also refers to the alkyl (as benzoyl) that comprises that one or more hydrogen atom is replaced by Sauerstoffatom, or by halogen, and better halogen is that the alkyl that replaces of fluorine is (as CF
3-or CF
3CH
2-).
A cyclic alkyl represented in term " cycloalkyl ".The term cycloalkyl also refers to comprise the cycloalkyl that contains at least one unsaturated group.The cycloalkyl example that is suitable for comprises cyclopropyl, cyclobutyl, cyclohexenyl, cyclohexadienyl and cyclohexyl.
Term " thiazolinyl " and " alkynyl " representative contain the alkyl (as allyl group, acetylene, ethene) of at least one unsaturated group.
Term " aryl " representative contains the isocyclic part of at least one benzene type ring, can is selected from following group and replace by one or more: halogen, nitro, nitroso-group, SO
3R
12, PO
3RcRd, CONR
13R
14, C
1-6Alkyl, C
2-6Thiazolinyl, C
2-6Alkynyl, C
6-12Aralkyl, C
6-12Aryl, C
1-6Alkoxyl group, C
2-6Alkene oxygen base, C
2-6Alkynyloxy group, C
6-12Aryloxy, C (O) C
1-6Alkyl, C (O) C
2-6Thiazolinyl, C (O) C
2-6Alkynyl, C (O) C
6-12Aryl, C (O) C
6-12Aralkyl, C
3-10Heterocycle, hydroxyl, NR
13R
14, C (O) OR
12, cyano group, azido-, amidino groups or guanidine radicals;
Wherein, R
12, Rc, Rd, R
13And R
14Be selected from independently of one another: H, C
1-12Alkyl, C
2-12Thiazolinyl, C
2-12Alkynyl, C
6-14Aryl, C
3-12Heterocycle, C
3-18Heteroaralkyl, C
6-18Aralkyl;
Or Rc and Rd form a 5-10 unit heterocycle with oxygen;
Or R
13And R
14Form a 3-10 unit heterocycle with nitrogen.The aryl example comprises phenyl and naphthyl.
Term " aralkyl " representative is by C
1-6Alkyl, C
1-6Thiazolinyl or C
1-6Alkynyl is connected in the aryl (as, benzyl) of adjacent atom.
A saturated or unsaturated loop section represented in term " heterocycle ", is inserted with at least one heteroatoms (as oxygen, sulphur or nitrogen) in the described loop section, can choose wantonly by following groups to replace: halogen, nitro, nitroso-group, SO
3R
12PO
3RcRd, CONR
13R
14C
1-6Alkyl, C
2-6Thiazolinyl, C
2-6Alkynyl, C
6-12Aralkyl, C
6-12Aryl, C
1-6Alkoxyl group, C
2-6Alkene oxygen base, C
2-6Alkynyloxy group, C
6-12Aryloxy, C (O) C
1-6Alkyl, C (O) C
2-6Thiazolinyl, C (O) C
2-6Alkynyl, C (O) C
6-12Aryl, C (O) C
6-12Aralkyl, C
3-10Heterocycle, hydroxyl, N R
13R
14, C (O) OR
12Cyano group, azido-, amidino groups or guanidine radicals;
R wherein
12Rc, Rd, R
13And R
14Be selected from independently of one another: H, C1-12 alkyl, C
2-12Thiazolinyl, C
2-12Alkynyl, C
6-14Aryl, C
3-12Heterocycle, C
3-18Heteroaralkyl, C
6-18Aralkyl;
Or Rc and Rd form a 5-10 unit heterocycle with oxygen;
Or R
13And R
14Form a 3-10 unit heterocycle with nitrogen.
Should be understood that the term heterocycle represents monocycle or many rings (as, two rings).The heterocyclic example includes but not limited to: epoxide; Furans; Cumarone; Isobenzofuran; Oxathiolane; Dithiolane; Dioxolane; The pyrroles; Tetramethyleneimine; Imidazoles; Pyridine; Pyrimidine; Indoles; Piperidines; Morpholine; Thiophene and thiomorpholine (thiomorpholine).
Term " heteroaralkyl " representative is by C
1-6Alkyl, C
1-6Thiazolinyl or C
1-6Alkynyl is connected in the heterocyclic radical of adjacent atom.
When having sulphur atom, this sulphur atom can be different oxidation state, i.e. S, SO or SO
2All such oxidation state are all within the scope of the invention.
When having nitrogen-atoms, this nitrogen-atoms can be different oxidation state, i.e. N or NO.All such oxidation state are all within the scope of the invention.
Term " independently " refers to that a substituting group has identical or different definition to each.
Should be understood that not only difference of amount that The compounds of this invention is used for the treatment of, and with giving approach, need the disease character of treatment and patient's age with the patient's condition and different, finally by attending doctor or animal doctor's judgement with selected particular compound.Yet suitable dose better 0.5-60mg/kg/ days, is preferably in 1-20mg/kg/ days generally in about 0.1-750mg/kg body weight/day scope.
Required dosage provides with single dose usually, or provides with appropriate intervals with divided dose, as every day twice, three time, and four times or more times dosage.
Described compound gives with unit dosage form usually; For example the per unit formulation contains 10-1500mg, common 20-1000mg, the most frequently used 50-700mg active ingredient.
Ideally, give the peak plasma concentrations that active ingredient should be able to reach this active compound, about 1-75 μ M, better about 2-50 μ M, preferably about 3-30 μ M.This can pass through for example optional salt brine solution of intravenous injection 0.1-5% active ingredient, or the oral pill that contains about 1-500mg active ingredient reaches.Desired blood concentration can provide keep in about 0.01-5.0mg/kg/ hour by continuous transfusion, or kept by the intermittent infusion that contains about 0.4-15mg/kg active ingredient.
When The compounds of this invention or its pharmacy acceptable salt with unite when using the amount the when dosage of each compound can be same or different from this compound of independent use at second therapeutical agent of same virus.Suitable dose does not embarrass those skilled in the art to understand.
When being used for the treatment of,, preferably provide active ingredient with pharmaceutical composition although The compounds of this invention can give by starting compound.Therefore, the present invention also provides a kind of pharmaceutical composition, and it comprises The compounds of this invention or its pharmaceutically acceptable derivates, and one or more pharmaceutically acceptable carriers and optional other therapeutic and/or preventative component.Carrier must be " acceptable ", promptly with other component compatibility of preparation, and can not damage its recipient.
Pharmaceutical composition comprise be fit to oral, rectum, nose, local (comprising cheek and hypogloeeis), through skin, those compositions that vagina or non-enteron aisle (comprising intramuscular injection, subcutaneous and vein) give, or be suitable for sucking or being blown into those compositions of form of medication.If suitable, the method that can adopt pharmaceutical field all to know is prepared into dose unit separately.These methods comprise that the solid carrier that makes active compound and liquid vehicle or fine dispersion or both combine, if make product be configured as required preparation if desired.
Suitable pharmaceutical composition for oral administration normally exists with discontinuous unit, and as capsule, cachets or tablet respectively contain the active ingredient of predetermined amount; As pulvis or particle; As solution, suspension or as emulsion.Active ingredient can also be as pill, paste or paste.The tablet and the capsule that are used for oral administration can contain vehicle commonly used, as binding agent, and weighting agent, lubricant, disintegrating agent or wetting agent.Tablet can carry out dressing according to the method that this area is all known.Oral liquid can be, for example water-based or oily suspensions, and solution, emulsion, syrup or elixir form, or can exist by dryed product, prepare with water or other suitable vehicles with preceding.Such liquid preparation can contain additive commonly used, as suspension agent, and emulsifying agent, non-aqueous vehicle (comprising edible oil) or sanitas.
The compounds of this invention can be mixed with for parenteral administration (as passing through injection, for example medicine group injects or infuses continuously), with ampoule, and the syringe of prefilled, the unit dosage of low capacity transfusion exists, or is contained in the multi-dose container that is added with sanitas.Said composition can adopt the suspension in oiliness or the preparation of water-based vehicle, solution or emulsion form, and can contain preparaton such as suspension agent, stablizer and/or dispersion agent.Perhaps, this active ingredient can be a powder type, can obtain by the aseptic separation of sterile solid, or obtains by freeze-drying solution, and use is preceding with suitable vehicle such as aseptic, the apirogen water preparation.
For the topical of epidermis, The compounds of this invention can be prepared becomes ointment, creme or lotion, or as percutaneous plaster.This percutaneous plaster can contain penetration enhancers such as Linaool, isothymol, thymol, Citric Acid, menthol and t-methyl allylphenol.For example available water-based of ointment and creme or oleaginous base are with suitable thickening material that adds and/or jelling agent preparation.Lotion can also contain one or more emulsifying agents, stablizer, dispersion agent, suspension agent, thickening material or tinting material usually with water-based or oleaginous base preparation.
The composition that is fit to oral cavity local medication comprises: lozenge, and it contains the active ingredient in the normally sucrose and gum arabic or tragakanta in flavoured base; Pastille, it contains the active ingredient just like gelatinum and glycerine or sucrose and gum arabic etc. in inert base; And mouth wass, it contains active ingredient in suitable liquid vehicle.
Carrier vehicle in the pharmaceutical composition of suitable rectal administration is a solid, and this composition is the suppository of unitary dose preferably.Suitable vehicle comprises the other materials that theobroma oil and this area are commonly used, and is conventional by mixed active compound and vehicle softening or fusing, then cooling and be shaped in mould and prepare suppository.
The composition that is fit to vagina administration can pesseulum, wadding, and creme, gel, paste, whipping agent or sprays provide, and said composition also contains suitable vehicle known in the art except that containing active ingredient.
When being used for intranasal administration, but The compounds of this invention can liquid spray or dispersion powder or the use of drops form.Drops can also contain one or more dispersion agents, chaotropic agent or suspension agent with water-based or the preparation of non-aqueous matrix.The liquid spray routine is carried through pressurized package.
For inhalation, The compounds of this invention is used insufflator usually, and atomizer or pressurized package are carried, or sends with the usual means of sending aerosol spray.Pressurized package can comprise suitable propelling agent, as Refrigerant 12, and trichlorofluoromethane, dichloro tetrafluoro ethane, carbonic acid gas or other suitable gas.With regard to pressurized aerosol, can send the dose unit of pre-metering by a valve.
Perhaps, for sucking or being blown into administration, The compounds of this invention can be taked the dry powder composite form, for example described compound and a kind of suitable powder matrix such as the powdered mixture of lactose or starch.Described dust composition can unit dosage exists, and for example, is contained in the capsule or in the tube, or as gelatin or send out blister and pack, give described powder by sucker or insufflator.
When needing, can adopt above-mentioned preparation to continue to discharge active ingredient.
Following general scheme and embodiment are provided, various embodiment of the present invention is described, but do not constitute limitation of the scope of the invention.
Embodiment 1
3-{[(2-carboxyl-5-phenyl-benzene sulphur-3-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-methyl }-piperidines, trifluoroacetate, compound 1.
Step I
(0.268g, 1.15mmol) (0.284g, (17mg 0.057mmol) handles THF 1.15mmol) (0.5mL) suspension with the dichloride dibutyl tin with 3-formyl radical N-Cbz-piperidines with 3-amino-5-phenyl-thiophene-2-carboxylic acid methyl esters.After 5 minutes, and the adding phenyl silane (156 μ L, 1.26mmol), stirring at room mixture 6 days.Solvent evaporated then, resistates are used CH by purification by silica gel column chromatography
2Cl
2: hexane: EtOAc obtains 3-[(1-methyl-piperidines-3-ylmethyl as elutriant)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters, be a kind of oily matter (0.2723g, 51% productive rate).
1H?NMR(CDCl
3，400MHz)：7.63-7.59(m，2H)，7.40-7.28(m，9H)，7.18-6.84(br?s，1H)，5.20(d，1H)，5.10(d，1H)，4.55(m，1H)，4.15(m，1H)，3.82(s，3H)，3.58-3.40(m，2H)，2.90(t，1H)，1.88-1,40(m，6H)。
Step II
With 3-[(1-methyl-piperidines-3-ylmethyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (162mg, 0.348mmol) be dissolved in 1, in the 2-ethylene dichloride (3.0mL), and with 1 of trans-4-methyl-cyclohexyl alkane carbonyl chloride, 2-ethylene dichloride (1.0mL, 0.43mmol) solution-treated.This solution of reflux 1 day.Solvent evaporated then, resistates uses hexane: EtOAc as elutriant by purification by silica gel column chromatography, obtains 3-[(4-methyl-cyclohexyl alkyl carbonyl)-(1-methyl-piperidines-3-ylmethyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters, be oily matter (0.194 g, 95% productive rate).
1H NMR (CDCl
3, 400 MHz): rotational isomer 65/35:7.90 (s, 0.35H), 7.70 (d, 0.65 H), 7.62-7.10 (m, 10H), 5.20-5.00 (m, 2H), 4.70 (m, 0.35H), 4.60-4.40 (m, 0.65H), 4.12 (m, 1H), 3.82 (s, 3H), 3.52 (t, 0.65H), 3.20 (t, 0.35H), 2.70 (d, 0.65H), 2.52 (t, 0.35H), 1.90 (m, 1H), 1.80-1.20 (m, 13H), 1.00-0.85 (m, 1H), 0.76 (d, 3H), 0.64 (m, 2H).
Step II I
With 3-[(4-methyl-cyclohexyl alkyl carbonyl)-(1-methyl-piperidines-3-ylmethyl)-amino]-(162mg 0.27mmol) is dissolved in THF: MeOH: H to 5-phenyl-thiophene-2-carboxylic acid methyl esters
2O (3: 2: 1.2.8mL) in the mixture, and use LiOHH
2(35mg 0.81mmol) handles O.55 ℃ of heated solutions 3 hours.Remove and desolvate, resistates is acidified to pH4 with HCl.Use the EtOAc extraction product, with salt water washing organic layer, dry and evaporation obtains 3-[(4-methyl-cyclohexyl alkyl carbonyl)-(1-methyl-piperidines-3-ylmethyl)-amino]-5-phenyl-thiophene-2-carboxylic acid (146mg, 92% productive rate).
1H?NMR(CDCl
3，400MHz)：9.98(br?s，1H)，7.80(d，1H)，7.62(d，1H)，7.48-7.24(m，9H)，5.20-5.05(m，2H)，4.35-3.95(m，3H)，3.00(m，1H)，2.85-2.52(m，2H)，2.15(m，1H)，1.82-1.18(m，12H)，0.78(d，3H)，0.68(m，2H)。
Step IV
With 3-[(4-methyl-cyclohexyl alkyl carbonyl)-(1-methyl-piperidines-3-ylmethyl)-amino]-(145mg 0.25mmol) is dissolved in CH to 5-phenyl-thiophene-2-carboxylic acid
3Among the CN (2.5mL), 0 ℃ of cooling, and with TMSI (144mL 1.0mmol) handles.0 ℃ of reaction stirred 1 hour was stirring at room 3 hours.Remove and desolvate resistates HCl acidifying.Product extracts with EtOAc, with salt water washing organic layer, drying.Solvent evaporated then, resistates use earlier the reversed-phase HPLC purifying, subsequently by purification by silica gel column chromatography, use CH
2Cl
2: MeOH: AcOH obtains 3-{[(2-carboxyl-5-phenyl-benzene sulphur-3-yl as elutriant)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-methyl }-piperidines trifluoroacetate (compound 1) (91.6mg, 66% productive rate).
1H?NMR(DMSO-d
6，400MHz)：7.92(br?s，1H)，7.66(m，2H)，7.49(s，1H)，7.42(m，2H)，7.33(m，1H)，4.50(m，1H)，3.33(m，3H)，2.80(m，1H)，2.56(m，1H)，2.30(m，2H)，1.80-1.30(m，8H)，1.20(m，3H)，0.73(d，3H)，0.73-0.45(m，2H)。
Prepare following compound in a similar way: compound 2. compound 3. compounds, 4. compounds 6 and compounds 14.
Embodiment 2
Chlorination 4-{[(2-carboxyl-5-phenyl-benzene sulphur-3-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-methyl }-1-methyl-piperidines, compound 38.
Step I
With 4-{[(2-methoxycarbonyl-5-phenyl-benzene sulphur-3-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-methyl }-(190mg 0.32mmol) is dissolved in MeOH (3.2mL), at H to piperidines-1-benzyl carboxylate
2(30psi) (3.2mmol), AcOH (1) and 10%Pd/C (97mg) handle for 37% solution, 0.36mL with formaldehyde down.Stirring at room reactant 48 hours, mixture filters on diatomite.Evaporate this solution and obtain resistates, use purification by silica gel column chromatography, use CH
2Cl
2: MeOH obtains 3-[(4-methyl-cyclohexyl alkyl carbonyl as elutriant)-(1-methyl-piperidin-4-yl methyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters, be oily matter (46.5mg, 31% productive rate).
1H?NMR(CDCl
3，400MHz)：7.62(d，2H)，7.43(m，3H)，7.10(s，1H)，3.97(m，1H)，3.85(s，3H)，3.20(m，3H)，2.48(s，3H)，2.42(m，1H)，2.10(m，1H)，1.85(m，3H)，1.70-1,40(m，8H)，1.30(m，2H)，0.78(d，3H)，0.68(m，2H)。
Step II
With 3-[(4-methyl-cyclohexyl alkyl carbonyl)-(1-methyl-piperidin-4-yl methyl)-amino]-(46mg 0.098mmol) is dissolved in THF: MeOH to 5-phenyl-thiophene-2-carboxylic acid methyl esters: H2O (3: 2: in mixture 1.1.0mL), and use LiOHH
2(12mg 0.29mmol) handles O.55 ℃ of heated solutions 3 hours.Remove and desolvate, resistates is acidified to pH 4 with HCl.Filtering precipitate with hexane wash and development, obtains chlorination 4-{[(2-carboxyl-5-phenyl-benzene sulphur-3-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-methyl }-1-methyl-piperidines (compound 38), be solid (35.3mg, 73% productive rate).
1H?NMR(CD
3OD，400MHz)：7.66(d，2H)，7.40(t，3H)，7.32(t，1H)，7.25(s，1H)，3.85(dd，1H)，3.52(dd，1H)，3.34(m，2H)，2.78(q，2H)，2.70(s，3H)，2.35(m，1H)，2.05(m，1H)，1.84(m，2H)，1.72(m，1H)，1.65-1.20(m，8H)，0.76(d，3H)，0.68(m，2H)。
Prepare following compound in a similar way: compound 33. compounds 35, compound 49 and compound 69.
Embodiment 3
The 3-[sec.-propyl-(the 5-methyl-[1,3] diox-2-carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid, compound 22.
The synthetic 5-methyl-[method of 1,3] diox-2-carboxylic acid: Tetrahedron (1989) 45, PP6987-6998.
Step I
At 0 ℃, the 5-methyl-[1,3] diox-2-carboxylic acid (and 53mg, 0.47mmol) 1,2-dichloroethane solution PPh
3(124mg, 0.47mmol), (63mg, 0.47mmol) (100mg 0.36mmol) handles NCS with sec.-propyl amino-5-phenyl-thiophene-2-carboxylic acid methyl esters.Reflux reactant 3 days.Evaporate this mixture and obtain resistates, use purification by silica gel column chromatography, use EtOAc: hexane is as elutriant, obtains 3-[sec.-propyl-(5-methyl-[1,3] diox-2-carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (51.8mg, 35% productive rate).
1H?NMR(CDCl
3，400MHz)：7.65(d，2H)，7.42(m，3H)，7.10(s，1H)，4.90(q，1H)，4.55(s，1H)，4.00(dd，1H)，3.90(dd，1H)，3.85(s，3H)，3.10(t，1H)，2.95(t，1H)，2.10(m，1H)，1.25(d，3H)，1.05(d，3H)，0.58(d，3H)。
Step II
With 3-[sec.-propyl-(5-methyl-[1,3] diox-2-carbonyl)-amino]-(49mg 0.12mmol) is dissolved in THF: MeOH: H to 5-phenyl-thiophene-2-carboxylic acid methyl esters
2O (3: 2: in mixture 1.1.1mL), use LiOHH
2(14mg 0.36mmol) handles O.55 ℃ of heated solutions 3 hours.Remove and desolvate, resistates is acidified to pH 4 with HCl.Product extracts with EtOAc, and with salt water washing organic layer, dry and evaporation obtains 3-[sec.-propyl-(5-methyl-[1,3] diox-2-carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid (compound 22), be solid (46.7mg, 98% productive rate).
1H?NMR(CD
3OD，400MHz)：7.65(d，2H)，7.45(m，3H)，7.30(s，1H)，4.80(q，1H)，4.80(s，1H)，4.00(dd，1H)，3.88(dd，1H)，3.08(t，1H)，2.98(t，1H)，2.00(m，1H)，1.30(d，3H)，1.05(d，3H)，0.60(d，3H)。
Embodiment 4
5-(3-fluoro-phenyl)-3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-thiophene-2-carboxylic acid compound 51.
Step I
With 3-amino-5-bromo-thiophene-2-carboxylic acid methyl esters (1.03g, 4.38mmol) anhydrous THF (1.1ml) suspension with 1,4-cyclohexanedione list ethene ketal (cyclohexanedione monoethyleneketal) (684mg, 4.38mmol) handle, (133mg 0.44mmol) handles to use the dichloride dibutyl tin subsequently.After 5 minutes, (877 μ l, 4.8mmol), this reaction mixture of stirring at room 2 days produces clear solution to add phenyl silane.Concentrate this solution then, the resistates purification by silica gel column chromatography, use EtOAc: hexane is as elutriant, obtains 5-bromo-3-(1,4-dioxa-spiral shell [4.5] last of the ten Heavenly stems-8-base amino)-thiophene-2-carboxylic acid methyl esters (1.11g, 68% productive rate).
1H?NMR(CDCl
3，400MHz)：6.90(br?s，1H)，6.65(s，1H)，3.95(s，4H)，3.78(s，3H)，3.35(m，1H)，2.00(m，2H)，1.80(m，2H)，1.65(m，4H)。
Step II
At 0 ℃, with trans 4-methyl-cyclohexyl alkane carboxylic acid (0.629g, 4.42mmol) 1,2-ethylene dichloride (30ml) solution triphenylphosphine (1.16g, 4.42mmol), (0.59g is 4.42mmol) with 5-bromo-3-(1 for N-chloro-succinimide, 4-dioxa-spiral shell [4.5] last of the ten Heavenly stems-8-base is amino)-(1.10g 2.92mmol) handles the thiophene-2-carboxylic acid methyl esters.In 90 ℃ of mixtures that stir to form 36 hours, concentrate then.The resistates purification by silica gel column chromatography, use EtOAc: hexane is as elutriant, obtain required product, 5-bromo-3-[(1,4-dioxa-spiral shell [4.5] last of the ten Heavenly stems-8-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-thiophene-2-carboxylic acid methyl esters and corresponding ketone, 5-bromo-3-[(4-methyl-cyclohexyl alkyl carbonyl)-(4-oxygen-cyclohexyl)-amino]-1: 1 mixture (537mg) of thiophene-2-carboxylic acid methyl esters.
Step II I
With 5-bromo-3-[(1,4-dioxa-spiral shell [4.5] last of the ten Heavenly stems-8-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-thiophene-2-carboxylic acid methyl esters and 5-bromo-3-[(4-methyl-cyclohexyl alkyl carbonyl)-(4-oxygen-cyclohexyl)-amino]-mixture of thiophene-2-carboxylic acid methyl esters (352mg) is dissolved in the tetrahydrofuran (THF) (4ml), and (4ml) handles with 3NHCl solution.Stirring at room reactant 20 hours is used ethyl acetate (10ml) dilution then.Isolate organic layer, water ethyl acetate (2 * 10mL) washed twice.The ethyl acetate layer that merges is used Na with salt solution (10ml) washing
2SO
4Drying is filtered and is concentrated.Resistates is by purification by silica gel column chromatography, and use EtOAc: hexane obtains 5-bromo-3-[(4-methyl-cyclohexyl alkyl carbonyl as elutriant)-(4-oxygen-cyclohexyl)-amino]-the thiophene-2-carboxylic acid methyl esters, be solid (296 mg).
1HNMR(CDCl
3，400MHz)：6.85(s，1H)，5.04(m，1H)，3.82(s，3H)，2.58-2.30(m，4H)，2.18(m，1H)，2.06(m，1H)，1.90(m，1H)，1.70-1.52(m，6H)，1,48-1.28(m，3H)，0.80(d，3H)，0.68(m，2H)。
Step IV
With 5-bromo-3-[(4-methyl-cyclohexyl alkyl carbonyl)-(4-oxygen-cyclohexyl)-amino]-(473mg 1.04mmol) is dissolved in the methyl alcohol (10.4ml) the thiophene-2-carboxylic acid methyl esters, is cooled to 0 ℃, and (43mg 1.14mmol) handles with sodium borohydride.0 ℃ was stirred this reactant after 30 minutes, stirring at room 30 minutes, and (20ml) goes out suddenly with 10% hydrochloric acid soln.(3 * 10mL) extract water, the dry ethyl acetate layer (Na that merges with ethyl acetate
2SO
4) and concentrate.The resistates purification by silica gel column chromatography, use EtOAc: hexane obtains 5-bromo-3-[(4-hydroxyl-cyclohexyl as elutriant)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-thiophene-2-carboxylic acid methyl esters (365mg, 77% productive rate), be solid.
1H?NMR(CDCl
3，400MHz)：6.82(s，1H)，4.56(m，1H)，3.82(s，3H)，3.45(m，1H)，2.08-1.72(m，4H)，1.75(m，1H)，1.68-1.23(m，11H)，0.98(m，1H)，0.80(d，3H)，0.68(m，2H)。
Step V
With 5-bromo-3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-(70mg, 0.15mmol) (32mg is 0.23mmol) at DME (2.0mL) and 2M Na with 3-fluorophenyl boric acid for the thiophene-2-carboxylic acid methyl esters
2CO
3De-gassed solution in the mixture of the aqueous solution (1.0mL) Pd (PPh
3)
4(17.6mg 0.015mmol) handles.Reflux reactant 18 hours.Reaction mixture dilutes with ethyl acetate and water.Separate organic layer, use the salt water washing, drying and concentration residue, use the preparative chromatography purifying, use EtOAc: hexane is as elutriant, obtain 5-(3-fluoro-phenyl)-3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkane carbonyl)-amino]-the thiophene-2-carboxylic acid methyl esters, be oily matter (61.7mg) that polluting has inexpungible triphenylphosphine oxide.
1H?NMR(CDCl
3，400MHz)：7.65(dd，3H)，7.53(t，2H)，7.43(m，5H)，7.32(m，1H)，7.1(m，1H)，7.02(s，1H)，4.56(m，1H)，3.82(s，3H)，3.40(m，1H)，2.14(br?s，1H)，2.05-1.88(m，4H)，1.78(m，1H)，1.68-1.54(m，5H)，1.51-1.26(m，4H)，0.98(m，1H)，0.75(d，3H)，0.72-0.54(m，2H)。
Step VI
With 5-(3-fluoro-phenyl)-3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-(61mg 0.13mmol) is dissolved in 4: 1 De diox: H to the thiophene-2-carboxylic acid methyl esters
2In the mixture of O (1.3 ml), and use LiOHH
2(20mg 0.476mmol) handles O.55 ℃ were stirred after 3 hours, removed and desolvated, and were distributed in the H that 5ml is acidified to pH 4 then
2Between O and the 5ml EtOAc.Separate organic layer, water ethyl acetate (2 * 5mL) washed twice.With the ethyl acetate layer drying (Na that merges
2SO
4) and concentrate.Resistates is by preparation type chromatography purification (10%MeOH/CH
2Cl
2), obtain 5-(3-fluoro-phenyl)-3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-thiophene-2-carboxylic acid (compound 51), be white solid (20.1mg, 34% productive rate).
1H?NMR(DMSO-d
6，400MHz)：7.72(d，1H)，7.60(m，2H)，7.50(m，1H)，7.24(m，1H)，4.50(d，1H)，4.28(m，1H)，3.18(m，1H)，1.95(m，1H)，1.85-1.10(m，14H)，0.88(m，1H)，0.75(d，3H)，0.68-0.45(m，2H)。
Prepare following compound in a similar way: compound 50. compounds 59, compound 60 and compound 61.
Embodiment 5
3-[(1-methylamino formyl radical-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid compound 71.
Step I
With 3-[(4-methyl-cyclohexyl alkyl carbonyl)-piperidin-4-yl-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (145mg, CH 0.33mmol)
2Cl
2(3.3mL) solution Et
3N (69mL, 0.49mmol) and methyl isocyanate (28.2mg 0.49mmol) handles.After the stirring at room 18 hours, keep initial substance.In this reactant, add methyl isocyanate (28.2mg, 0.49mmol), restir 4 hours.Solvent evaporated is dissolved in resistates among the EtOAc, with HCl (0.1M) and salt water washing, and dry (Na
2SO
4) and concentrate.Resistates uses (5%MeOH/CH by purification by silica gel column chromatography
2Cl
2), obtain 3-[(1-methylamino formyl radical-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (142mg, 87% productive rate).
1H?NMR(CDCl
3，400MHz)：7.65(m，2H)，7.45(m，3H)，7.00(s，1H)，4.78(m，1H)，4.35(q，1H)，4.05(dd，1H)，3.90(m，1H)，3.85(s，3H)，2.85(m，2H)，2.75(d，3H)，1.95(m，2H)，1.80(m，1H)，1.70-1.55(m，5H)，1.50-1.25(m，3H)，1.10(m，1H)，0.78(d，3H)，0.75-0.58(m，2H)。
Step II
With 3-[(1-methylamino formyl radical-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-(139mg 0.28mmol) is dissolved in 4: 1 De diox: H to 5-phenyl-thiophene-2-carboxylic acid methyl esters
2In the O mixture (2.8ml), use LiOHH
2(35.3mg 0.84mmol) handles O.55 ℃ were stirred after 3 hours, removed and desolvated, and were distributed in the H that 5ml is acidified to pH 4 then
2Between O and the 5ml EtOAc.Separate organic layer, water ethyl acetate (2 * 5mL) washed twice.With the ethyl acetate layer drying (Na that merges
2SO
4) and concentrate.Resistates uses (10%MeOH/CH by purification by silica gel column chromatography
2Cl
2), obtain 3-[(1-methylamino formyl radical-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid (compound 71), be white solid (101.8mg, 75% productive rate).
1H?NMR(CD
3OD，400MHz)：7.72(m，2H)，7.47-7.37(m，3H)，7.32(s，1H)，4.65(m，1H)，4.00(m，1H)，2.82(q，2H)，2.65(d，3H)，2.10(m，1H)，1.90(m，2H)，1.80-1.22(m，8H)，1.14(m，1H)，0.78(d，3H)，0.75-0.55(m，2H)。
Embodiment 6
3-[[1,3] dioxolane-2-ylmethyl-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid compound 8.
Step I
Adopt the Pd coupling method described in the embodiment 32, preparation 3-[([1,3] dioxolane-2-ylmethyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters.
Step II
3-[[1,3] dioxolane-2-ylmethyl-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters, adopt the method described in the embodiment 32 that is similar to.
Step II I 3-[[1,3] dioxolane-2-ylmethyl-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid, adopt the method described in the embodiment 32 that is similar to.
Adopt similar approach to prepare compound 7.
Embodiment 7
5-(3-fluoro-phenyl)-3-[(2-hydroxy-4-methyl-hexanaphthene carbonyl)-sec.-propyl-amino]-thiophene-2-carboxylic acid compound 44.
Step I
(210mg, 0.755mmol) (140mg is 0.116mmol) at DME (8mL) and 2M Na with 3-fluorophenyl boric acid at 5-bromo-3-sec.-propyl amino-2-carboxylate methyl ester
2CO
3The de-gassed solution of mixture (4mL) in add Pd (PPh
3)
4(43mg), under refluxad, at N
2Stirred reaction mixture is 3 hours under the atmosphere.Reaction mixture dilutes with ethyl acetate and water.Separate organic layer, dry (Na
2SO
4) and concentrate.Separate 5-(3-fluoro-the phenyl)-3-sec.-propyl amino-thiophene-2-carboxylic acid methyl esters (200mg, 91%) that obtains molasse.
1H?NMR(CDCl
3，400MHz)：δ7.50-7.25(m，3H)，7.13-7.05(m，1H)，6.88(s，1H)，6.74(bs，1H)，3.80(s，3H)，3.75(m，1H)，1.35，1.30(2s，6H)。
Step II
At N
2Under the atmosphere, to 5-(3-fluoro-phenyl)-3-sec.-propyl amino-thiophene-2-carboxylic acid methyl esters (200mg, 0.683mmol) 1, add in 2-ethylene dichloride (5mL) solution, acetate 2-is chloroformyl-5-methyl-cyclohexyl base ester (148mg, 0.679mmol) and triphenylphosphine (197mg, 0.751mmol).Reaction mixture refluxed 12 hours is then with chloroform and water dilution.Separate organic layer, dry (Na
2SO
4) and concentrate.Resistates uses the hexane solution purifying of 15% ethyl acetate with preparation type TLC plate, obtains 3[(2-acetoxyl group-4-methyl-cyclohexyl alkyl carbonyl)-sec.-propyl-amino]-5-(3-fluoro-phenyl)-thiophene-2-carboxylic acid methyl esters, be white solid (40mg, 12%).
1HNMR(CDCl
3，400MHz)：δ7.45-7.25(m，4H)，7.13-6.95(m，1H)，5.13(m，1H)，4.87-4.75(m，1H)，3.80(s，3H)，2.37-0.62(m，20H)。
Step II I
3[(2-acetoxyl group-4-methyl-cyclohexyl alkyl carbonyl)-sec.-propyl-amino]-(30mg 0.063mmol) adds THF: MeOH: H to 5-(3-fluoro-phenyl)-thiophene-2-carboxylic acid methyl esters
2O (3: 2: 1.2mL) in the mixture, add 1N LiOHH then
2The O aqueous solution (0.38mL, 0.380mmol).Stirring at room reaction mixture 12 hours.Remove and desolvate, resistates is distributed between water and the ethyl acetate.Waterbearing stratum 10%KHSO
4The solution acidifying.Separate organic layer, dry (Na
2SO
4) and concentrate.The resistates purification by silica gel column chromatography, use chloroform and methyl alcohol (8: 2), obtain 5-(3-fluoro-phenyl)-3-[(2-hydroxy-4-methyl-hexanaphthene carbonyl)-sec.-propyl-amino]-thiophene-2-carboxylic acid (compound 44) (15mg, 58%), be white solid, two kinds of rotational isomers are arranged.
1HNMR(CDCl
3，400MHz)：δ7.50-7.25(m，3H)，7.06(m，2H)，6.25(bs，1H)，5.25(s，1H，minor)，4.87(s，1H，major)，4.13(s，1H，major)，3.87(s，1H，minor)，2.38-0.45(m，17H)。ESI(M-H)：418。
Prepare compound 62 in a similar way.
Embodiment 8
3-[sec.-propyl-(4-methylene radical-hexanaphthene carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid compound 28.
Step I
3-sec.-propyl amino-5-phenyl-thiophene-2-carboxylic acid methyl esters (1.5g, 5.45mmol) 1, add N-chlorine succinic diamide (0.940g in the 2-dichloroethane solution, 7.091mmol), triphenylphosphine (1.9g, 7.091mmol) and 4-oxygen-hexahydrobenzoic acid (800mg, 5.455mmol).At N
2Under the atmosphere, the backflow stirred reaction mixture spends the night.Reaction mixture CH
2Cl
2NaHCO is used in dilution
3The saturated solution extraction.Separate organic layer, dry (Na
2SO
4) and concentrate.The resistates silica gel column chromatography, use ethyl acetate: hexane (1: 4) obtains 3-[sec.-propyl-(4-oxygen-hexanaphthene carbonyl)-amino as the elutriant purifying]-5-phenyl-thiophene-2-carboxylic acid methyl esters (1.2g, 55%), be syrup.
1H?NMR(CDCl
3，400MHz)：δ7.75-7.50(m，2H)，7.70-7.38(m，3H)，7.12(s，1H)，4.95(m，1H)，3.87(s，3H)，2.75-0.83(m，15H)。
Step II
((939mg is in THF 2.630mmol) (10mL) cold soln (78 ℃) 2.280mmol) to join the Diethylaminoethyl triphenyl phosphonium for 2.5M, 0.9mL with butyllithium.Stirring at room reaction mixture 1 hour adds 3-[sec.-propyl-(4-oxygen-hexanaphthene carbonyl)-amino in-78 ℃]-THF (5mL) solution of 5-phenyl-thiophene-2-carboxylic acid methyl esters (700mg, 1.754).Stirring at room reaction mixture 12 hours.By adding NH
4The Cl saturated solution goes out reaction suddenly, dilutes with ethyl acetate.Separate organic layer, dry (Na
2SO
4) and concentrate.Resistates uses ethyl acetate by purification by silica gel column chromatography: hexane (1: 4) obtains 3-[sec.-propyl-(4-methylene radical-hexanaphthene carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (300mg, 43%), be solid.
1HNMR(CDCl
3，400MHz)：δ7.63(d，2H)，7.50-7.38(m，3H)，7.12(s，1H)，4.99(m，1H)，4.55(d，2H)，3.85(s，3H)，2.25(m，3H)，1.83-1.63(m，5H)，1.50(m，1H)，1.25(d，3H)，0.99(d，3H)。
Step II I
With 3-[sec.-propyl-(4-methylene radical-hexanaphthene carbonyl)-amino]-(50mg 0.126mmol) adds THF: MeOH: H to 5-phenyl-thiophene-2-carboxylic acid methyl esters
2O (3: 2: 1, in mixture 3mL), add 1N LiOH H then
2The O aqueous solution (0.8mL, 0.800mmol).Stirring at room reaction mixture 12 hours.Remove and desolvate, resistates is distributed between water and the ethyl acetate.Waterbearing stratum 10%KHSO
4The solution acidifying.Separate organic layer, dry (Na
2SO
4) and concentrate.Resistates uses chloroform and methyl alcohol (8: 2) by purification by silica gel column chromatography, obtains 3-[sec.-propyl-(4-methylene radical-hexanaphthene carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid (compound 28) (25mg, 52%), be white solid.
1H?NMR(CDCl
3，400MHz)：δ7.61(d，J＝7Hz，2H)，7.40-7.38(m，3H)，7.04(s，1H)，4.92(m，1H)，4.50(d，J＝7.6Hz，2H)，2.21-1,43(m，9H)，1.15(bd，3H)，0.93(bd，3H)。
Embodiment 9
5-(4-fluoro-phenyl)-3-[sec.-propyl-(4-methyl-cyclohexyl-3-alkene carbonyl)-amino]-thiophene-2-carboxylic acid compound 53.
Step I
3-amino-5-(4-fluoro-phenyl)-thiophene-2-carboxylic acid methyl esters (500mg, 2.0mmol) 1, add successively in the stirred solution of 2-ethylene dichloride (10mL) 2-methoxyl group propylene (0.76mL, 8.0mmol), AcOH (0.12mL, 4.0mmol) and NaBH (OAc)
3(0.848mg 8.0mmol), stirred 16 hours.Use EtOAc and H then
2The O dilution.Add NaHCO
3This aqueous solution is adjusted to pH=7.Water extracts with EtOAc, and MgSO is used in the salt water washing of the extraction liquid of merging
4Drying is filtered.With hexane to the 5%EtOAc-hexane carry out purifying in conjunction with elutriant, obtain 5-(4-fluoro-phenyl)-3-sec.-propyl amino-thiophene-2-carboxylic acid methyl esters (0.530mg, 91% productive rate).
1H?NMR(CDCl
3，400MHz)：δ7.62(d，2H)，7.09(m，2H)，6.81(s，1H)，3.84(s，3H)，3.71(m，1H)，1.35(d，6H)。
Step II
According to Journal of Organic Chemistry (1986) 51 (23), PP4485-8; Middle reported method prepares 4-methyl-cyclohexyl-3-alkene dicarbonyl chloride.With this 4-methyl-cyclohexyl-3-alkene dicarbonyl chloride (0.121g, 0.77mmol) with 5-(4-fluoro-phenyl)-3-sec.-propyl amino-thiophene-2-carboxylic acid methyl esters (0.150g, 0.51mmol) be dissolved in together anhydrous 1, in the 2-ethylene dichloride (2mL).Backflow stirred reaction mixture 16 hours.Then, remove and to desolvate, resistates by the flash chromatography purifying (8: 2 hexanes/EtOAc), obtain 140mg (66%) 5-(4-fluoro-phenyl)-3-[sec.-propyl-(4-methyl-cyclohexyl-3-alkene carbonyl)-amino]-the thiophene-2-carboxylic acid methyl esters.
1HNMR(CDCl
3?400MHz)：δ7.60(m，2H)，7.15(m，2H)，7.02(d，1H)，5.42-5.20(m，1H)，4.99(m，1H)，3.83(d，3H)，2.41-1.50(m，10H)，1.20(m，3H)，0.98(d，3H)。
Step II I
With 5-(4-fluoro-phenyl)-3-[sec.-propyl-(4-methyl-cyclohexyl-3-alkene carbonyl)-amino]-(0.140g 0.34mmol) adds THF: MeOH: H to the thiophene-2-carboxylic acid methyl esters
2O (3: 2: in mixture 1.10mL), add 1N LiOHH then
2The O aqueous solution (2.1mL, 2.04mmol).50 ℃ of stirred reaction mixtures 1 hour.Remove and desolvate, resistates is distributed between water and the ethyl acetate.Waterbearing stratum 10%KHSO
4The solution acidifying.Separate organic layer, dry (Na2SO4) also concentrates.Resistates uses methylene dichloride: methyl alcohol (9: 1) purifying, acquisition 5-(4-fluoro-phenyl)-3-[sec.-propyl-(4-methyl-cyclohexyl-3-alkene carbonyl)-amino with preparation type TLC]-thiophene-2-carboxylic acid (compound 53) (31mg, 23%).
1H?NMR(CDCl
3，400MHz)：δ7.81(m，2H)，7.43(d，1H)，7.28(m，2H)，5.38-5.16(m，1H)，4.72(m，1H)，2.20(d，2H)，1.95-1.20(m，8H)，1.12(m，3H)，0.90(d，3H)。
Prepare following compound in a comparable manner: compound 17.
Embodiment 10
Trans-3-[(1-fluoro-4-methyl-cyclohexyl alkyl carbonyl)-sec.-propyl-amino]-5-phenyl-thiophene-2-carboxylic acid compound 9.
Step I
At 3-amino-5-phenyl-thiophene-2-carboxylic acid methyl esters (1.82g, 7.8mmol) 1, order adds 2-methoxyl group propylene (3.0mL in the stirred solution of 2-ethylene dichloride (40mL), 31.2mmol), AcOH (1.8mL, 31.2mmol) and NaBH (OAc) 3 (3.31g 15.6mmol), stirred 2 hours.Use EtOAc and H then
2The O dilution.Add NaHCO
3Regulate this solution to pH=7.Water extracts with EtOAc, with the extraction liquid salt water washing that merges, uses MgSO
4Drying is filtered.With hexane to the 5%EtOAc-hexane carry out purifying in conjunction with elutriant, obtain 3-sec.-propyl amino-5-phenyl-thiophene-2-carboxylic acid methyl esters (2.07g, 96% productive rate).
1H?NMR(CDCl
3，400MHz)：δ7.62(d，2H)，7.40(m，3H)，6.91(s，1H)，3.84(s，3H)，3.71(m，1H)，1.35(d，6H)。
Step II
According to Synthesis, reported method prepares suitable/anti-form-1-fluoro-4-methyl-cyclohexyl alkane carboxylic acid among April (1998) PP310-313.At 0 ℃, should suitable/anti-form-1-fluoro-4-methyl-cyclohexyl alkane carboxylic acid (0.220g, 1.37mmol) and PPh
3(0.360g, 1.37mmol) be dissolved in together anhydrous 1, in the 2-ethylene dichloride (20mL).Add then NCS (0.181g, 1.37mmol) and 3-sec.-propyl amino-5-phenyl-thiophene-2-carboxylic acid methyl esters (0.290g, 1.05mmol), backflow stirred reaction mixture 16 hours.After being cooled to room temperature, the saturated NaHCO of crude product
3Washing.Organic layer drying (MgSO4) concentrates, and resistates is with preparation type TLC plate layer chromatography purifying (20%EtOAc/ hexane), obtain 171mg (39%) suitable/trans 3-[(1-fluoro-4-methyl-cyclohexyl alkyl carbonyl)-sec.-propyl-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters.
1H?NMR(CDCl
3，400MHz)：majorδ6.61(d，2H)，6.40(m，3H)，7.03(s，1H)，4.93(m，1H)，3.81(s，3H)，2.18-1.30(m，7H)，1.20(d，3H)，1.10(m，2H)，0.96(d，3H)，0.81(d，3H)。
Step II I
3-[(1-fluoro-4-methyl-cyclohexyl alkyl carbonyl)-sec.-propyl-amino]-(0.049g 0.12mmol) adds THF: MeOH: H to 5-phenyl-thiophene-2-carboxylic acid methyl esters
2O (3: 2: 1.10mL) in the mixture, add 1N LiOHH then
2The O aqueous solution (0.35mL, 0.35mmol).50 ℃ of stirred reaction mixtures 3 hours.Remove and desolvate, resistates is distributed between water and the ethyl acetate.Waterbearing stratum 10%KHSO
4The solution acidifying.Separate organic layer, dry (Na
2SO
4) and concentrate.Resistates preparation type TLC, use methylene dichloride: methyl alcohol (9: 1) purifying obtains trans-3-[(1-fluoro-4-methyl-cyclohexyl alkyl carbonyl)-sec.-propyl-amino]-5-phenyl-thiophene-2-carboxylic acid (compound 9) (9mg, 19%).
1H?NME(MeOD，400MHz)：δ6.75(d，2H)，6.41(m，3H)，7.29(s，1H)，4.85(m，1H)，2.1-1.85(m，4H)，1.59-1.24(m，3H)，1.22(d，3H)，1.10(m，2H)，0.99(d，3H)，0.81(d，3H)。
Prepare following compound in a comparable manner: compound 10.
Embodiment 11
3-[N-(2,4-two chloro-benzoyls)-N ', N '-dimethyl-diazanyl]-5-phenyl-thiophene-2-carboxylic acid compound 72.
Step I
At 3-bromo-5-phenyl-thiophene-2-carboxylic acid methyl esters (0.500g, 1.68mmol) toluene (10ml) solution in add N, N-dimethyl-hydrazine (0.121g, 2.02mmol), cesium carbonate (0.767g, 2.36mmol), BINAP (0.106g, 0.17mmol) and acid chloride (II) (0.019g, 0.08mmol).110 ℃ of stirred reaction mixtures 16 hours.Mixture is distributed between toluene (20mL) and the water (20mL), separates organic layer.(2 * 10mL) washed twice are with the toluene layer drying (MgSO that merges with toluene for water
4), concentrate, resistates preparation type TLC (10%EtOAc/ hexane) purifying, the 3-of acquisition 0.350g (75%) (N ', N '-dimethyl-diazanyl)-5-phenyl-thiophene-2-carboxylic acid methyl esters.NMR
1H(CDCl
3，400MHz)：δ7.71(d，2H)，7.40(m，3H)，7.13(s，1H)，3.87(s，3H)，2.65(s，6H)。
Step II
At N
2Under the atmosphere, 3-(N ', N '-dimethyl-diazanyl)-5-phenyl-thiophene-2-carboxylic acid methyl esters (0.200g, 0.72mmol) 1, add 2 in 2-ethylene dichloride (10ml) solution, 4-two chloro-Benzoyl chlorides (0.228g, 1.08mmol).Backflow stirred reaction mixture 1.5 hours.Then, remove and to desolvate, resistates with preparation type TLC purifying (8: 2 hexanes/EtOAc), obtain 0.017g (5%) 3-[N-(2,4-two chloro-benzoyls)-N ', N '-dimethyl-diazanyl]-5-phenyl-thiophene-2-carboxylic acid methyl esters.NMR
1H(CDCl
3，400MHz)：δ7.62(m，2H)，7.40(m，3H)，7.23(d，1H)，3.87(s，3H)，2.52(s，6H)。
Step II I
3-[N-(2,4-two chloro-benzoyls)-N ', N '-dimethyl-diazanyl]-(0.050g 0.11mmol) adds THF: MeOH: H to 5-phenyl-thiophene-2-carboxylic acid methyl esters
2O (3: 2: 1.10mL) in the mixture, add 1N LiOHH then
2The O aqueous solution (0.67mL, 0.67mmol).60 ℃ of stirred reaction mixtures 2 hours.Remove and desolvate, resistates is distributed between water and the ethyl acetate.Waterbearing stratum 10%KHSO
4The solution acidifying.Separate organic layer, dry (Na
2SO
4) and concentrate.Resistates uses methylene dichloride: methyl alcohol (9: 1) purifying, acquisition 3-[N-(2,4-two chloro-benzoyls)-N ', N '-dimethyl-diazanyl with preparation type TLC]-5-phenyl-thiophene-2-carboxylic acid (compound 72) (0.008g, 17%).
1H?NMR(DMSO，400MHz)δ7.81(d，2H)，7.69(d，2H)，7.54-7.40(m，5H)，2.42(s，6H)。
Prepare following compound in a comparable manner: compound 64.
Embodiment 12
5-(3-fluoro-phenyl)-3-[sec.-propyl-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-thiophene-2-carboxylic acid compound 5.
Step I
The 3-fluorobenzoic boric acid (25.0mg, 0.180mmol) and 5-bromo-3-[sec.-propyl-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-thiophene-2-carboxylic acid methyl esters (24mg, 0.060mmol) adding Pd (PPh in the mixture (1.0mL) of toluene/MeOH of 5: 1
3) 4 (toluene solution 10mol%) (0.5mL) adds 2M Na subsequently for 7.0mg, 0.006mmol
2CO
3The aqueous solution (0.06mL, 0.120mmol).The reaction mixture that forms was heated 18 hours for 70 ℃, be cooled to room temperature, pass through MgSO
4Filter, wash with EtOAc.Solvent evaporated, resistates are the elutriant purifying with ethyl acetate/hexane (20: 80) on preparation type TLC, obtain (25.0mg, 99% productive rate) 5-(3-fluoro-phenyl)-3-[sec.-propyl-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-the thiophene-2-carboxylic acid methyl esters.
1H?NMR(CDCl
3，400MHz)：7.45-7.39ppm(m，2H)；7.34-7.31ppm(m，1H)；7.13-7.07ppm(m，1H)；7.06ppm(s，1H)；5.00-4.93ppm(m，1H)；3.85ppm(s，3H)；2.04-1.95ppm(m，1H)；1.74-1.57ppm(m，5H)；1,48-1.38ppm(m，1H)；1.36-1.27ppm(m，1H)；1.17ppm(d，3H)；0.94ppm(d，3H)；0.77ppm(d，3H)；0.73-0.55ppm(m，2H)。
Step II
With 5-(3-fluoro-phenyl)-3-[sec.-propyl-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-(25mg 0.060mmol) is dissolved in 4: 1 De diox: H to the thiophene-2-carboxylic acid methyl esters
2In the O mixture (0.8mL), and adding LiOH1N (0.3ml, 0.300mml).After the stirring at room 3 hours, remove and desolvate, be distributed in the H that 10ml is acidified to pH4
2Between O and the 10ml EtOAc.Separate organic layer, water ethyl acetate (2 * 10mL) washed twice.With the ethyl acetate layer drying (Na that merges
2SO
4), concentrate resistates preparation type chromatography purification (10%MeOH/CH
2Cl
2), 5-(3-fluoro-phenyl)-3-[sec.-propyl-(4-methyl-cyclohexyl alkyl carbonyl)-amino of acquisition 20mg (83%)]-thiophene-2-carboxylic acid (compound 5).
1H?NMR(CD
3OD，400MHz)：7.57-7.44ppm(m，3H)；7.39ppm(s，1H)；7.17-7.11ppm(m，1H)；4.87-4.81ppm(m，1H)；2.15-2.09ppm(m，1H)；1.82-1.78ppm(m，1H)；1.71-1.52ppm(m，4H)；1,42-1.25ppm(m，2H)；1.22ppm(d，3H)；1.00ppm(d，3H)；0.78ppm(d，3H)；0.73-0.56ppm(m，2H)。
Embodiment 13
3-[(2-acetylamino-4-methyl-cyclohexyl alkyl carbonyl)-sec.-propyl-amino]-5-phenyl-thiophene-2-carboxylic acid compound 12.
Step I
With 2-hydroxy-4-methyl-hexahydrobenzoic acid ethyl ester (495mg; 2.66mmol) be dissolved among the THF (13ml); add then diphenylphosphine acyl group nitride (diphenylphosphoryl azide) (680 μ l, 3.19mmol) and triphenylphosphine (837mg, 3.19mmol).The solution that cooling forms in ice bath, and the adding diethylazodicarboxylate (502ul, 3.19mmol).After the stirring at room 20 hours, remove and desolvate, resistates obtains 365mg (65%) 2-azido--4-methyl-cyclohexyl alkane carboxylic acid, ethyl ester by flash chromatography purifying (0-3%EtOAc/ hexane).
Step II
(425mg 2.01mmol) is dissolved in 4: 1 De diox: H with 2-azido--4-methyl-cyclohexyl alkane carboxylic acid, ethyl ester
2In the O mixture (20ml), add then LiOH 1N (10ml, 10.05mmol).After the stirring at room 35 minutes, remove and desolvate, be distributed in the H that 15ml is acidified to pH4 then
2Between O and the 15ml EtOAc.Separate organic layer, water ethyl acetate (2 * 10mL) washed twice.Ethyl acetate layer drying (the Na that merges
2SO
4), concentrate, obtain 2-azido--4-methyl-cyclohexyl alkane carboxylic acid of 166mg and 2: 1 mixtures of 4-methyl-cyclohexyl-1-olefinic carboxylic acid.
Step II I
At 2-azido--4-methyl-cyclohexyl alkane carboxylic acid and 4-methyl-cyclohexyl-1-olefinic carboxylic acid (166mg, 0.91mmol) mixture of (2: 1) is in methylene dichloride (9ml) solution, (905 μ l 1.82mmol), add 1 dimethyl formamide subsequently to add 2.0M oxalyl chloride solution.Stirring at room reaction mixture 3 hours.Remove then and desolvate, obtain 2-azido--4-methyl-cyclohexyl alkane carboxylic acid chloride and 4-methyl-cyclohexyl-muriatic 2: 1 mixtures of 1-olefinic carboxylic acid of 182mg (99%).
Step IV
At 3-sec.-propyl amino-5-phenyl-thiophene-2-carboxylic acid methyl esters (227mg, 0.824mmol) 1, in 2-ethylene dichloride (2.5ml) solution, add and be dissolved in 1,2:12-azido-in the 2-ethylene dichloride (0.5ml)-4-methyl-cyclohexyl alkane carboxylic acid chloride and 4-methyl-cyclohexyl-muriatic mixture of 1-olefinic carboxylic acid (182mg, 0.906mmol).The solution of 90 ℃ of stirring formation 18 hours is cooled to room temperature then.With ethyl acetate (10ml) and saturated NaHCO
3Solution (10ml) dilution.Water phase separated is with ethyl acetate (2 * 10ml) washed twice, the organic layer drying (Na of merging
2SO
4), filter and concentrate.Resistates obtains 3-[(2-azido--4-methyl-cyclohexyl alkyl carbonyl of 178mg (49%) by flash chromatography purifying (0-20%EtOAc/ hexane))-sec.-propyl-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters.
Step V
At 3-[(2-azido--4-methyl-cyclohexyl alkyl carbonyl)-sec.-propyl-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (17.5mg, 0.04mmol) methyl alcohol (400 μ l) solution in add concentrated hydrochloric acid (15 μ l), add subsequently palladium on 10% gac (4mg, 0.004mmol).Feed 20psi hydrogen in the reactor, stirring at room is 17 hours then.Resistates leaches by diatomite, with methanol wash and evaporation, obtains crude product.In the crude product that is dissolved in methylene dichloride (400 μ l), add pyridine (19 μ l, 0.24mmol), add subsequently diacetyl oxide (15 μ l, 0.16mmol) and the DMAP of catalytic amount.The solution that stirring at room forms 24 hours is used NaHCO then
3Saturated solution (5ml) goes out suddenly.Water phase separated is with methylene dichloride (2 * 5ml) washings.With the organic layer drying (Na that merges
2SO
4), filter and concentrate.Resistates obtains 8.5mg (47%, two step) 3-[(2-acetylamino-4-methyl-cyclohexyl alkyl carbonyl by preparation type chromatography purification (60%EtOAc/ hexane))-sec.-propyl-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters.
Step VI
With 3-[(2-acetylamino-4-methyl-cyclohexyl alkyl carbonyl)-sec.-propyl-amino]-(8.5mg 0.019mmol) is dissolved in 4: 1 De diox: H to 5-phenyl-thiophene-2-carboxylic acid methyl esters
2In O (250ul) mixture, add then LiOH 1N (22 μ l, 0.023mmol).After the stirring at room 22 hours, remove and desolvate, be distributed in the H that 5ml is acidified to pH 4 then
2Between O and the 5ml EtOAc.Separate organic layer, water ethyl acetate (2 * 5mL) washed twice.With the ethyl acetate layer drying (Na that merges
2SO
4), concentrate, obtain 3-[(2-acetylamino-4-methyl-cyclohexyl alkyl carbonyl of 7.5mg (91%))-sec.-propyl-amino]-5-phenyl-thiophene-2-carboxylic acid (compound 12).
1H?NMR(CDCl
3，400MHz)7.78-7.73?ppm(m，2H)；7.60?ppm(s，1H)，7.49-7.39ppm(m，3H)；4.84-4.77?ppm(m，1H)；4.36-4.33?ppm(m，1H)；2.50-2.45?ppm(m，1H)；1.98?ppm(s?3H)；1.95-1.85?ppm(m，2H)；1.73-1,49?ppm(m，5H)；1.17?ppm(d，3H)；0.94?ppm(d，3H)；0.83-0.77?ppm(m?et?d，4H)。
Embodiment 13
3-[(4-methyl-cyclohexyl alkyl carbonyl)-(4-oxygen-cyclohexyl)-amino]-5-phenyl-thiophene-2-carboxylic acid compound 13.
Step I
Under the room temperature, (987mg adds 1 in anhydrous THF (1.0ml) solution 4.23mmol) at 3-amino-5-phenyl-thiophene-2-carboxylic acid methyl esters, 4-cyclohexanedione list ethene ketal (661mg, 4.23mmol), add subsequently the dichloride dibutyl tin (129mg, 0.42mmol).After 5 minutes, (575 μ L, 4.65mmol), stirring at room reaction mixture 4 days forms clear solution to add phenyl silane.Concentrate this solution, resistates is by flash chromatography purifying (0-30%EtOAc/ hexane), obtains 3-(1,4-Dioxa-spiral shell [4.5] last of the ten Heavenly stems-the 8-base is amino)-5-phenyl-thiophene-2-carboxylic acid methyl esters of 1.22g (77%).
1H?NMR(CDCl
3，400MHz)7.64-7.61?ppm(m，2H)；7.42-7.33?ppm(m，3H)；6.85ppm(s，1H)；3.96?ppm(s，4H)；3.82?ppm(s，3H)；3.49?ppm(bs，1H)；2.06-2.00ppm(m，2H)；1.85-1.81?ppm(m，2H)；1.79-1.63?ppm(m，4H)。
Step II
Trans 4-methyl-cyclohexyl alkane carboxylic acid (148mg, 1.044mmol) and triphenylphosphine (274mg 1.044mmol) is dissolved in 1, add in the solution of 2-ethylene dichloride (1.5ml) N-chloro-succinimide (145mg, 1.084mmol).After the stirring at room 15 minutes, add 1 of 3-(1,4-Dioxa-spiral shell [4.5] last of the ten Heavenly stems-the 8-base is amino)-5-phenyl-thiophene-2-carboxylic acid methyl esters (300mg, 0.803 ' mmol), 2-ethylene dichloride (1.5ml) solution.The mixture of 90 ℃ of stirring formation is 18 hours then, is cooled to room temperature then.With ethyl acetate (10ml) dilution, and add saturated NaHCO
3Solution (10ml).Water phase separated, and with ethyl acetate (2 * 10ml) washings are with the organic layer drying (Na that merges
2SO
4), filter and concentrate.Resistates obtains the 3-[(1 of 265mg (66%) by flash chromatography purifying (0-30%EtOAc/ hexane), 4-Dioxa-spiral shell [4.5] last of the ten Heavenly stems-8-yl)-(trans 4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters.
1H?NMR(CDCl
3，400MHz)7.66-7.61?ppm(m，2H)；7.47-7.38?ppm(m，3H)；7.04ppm(s，1H)；4.72-4.64?ppm(m，1H)；3.90-3.65?ppm(m，7H)；2.04-1.89?ppm(m，2H)；1.79-1.50?ppm(m，10H)；1,49-1.37?ppm(m，1H)；1.35-1.17?ppm(m，3H)；0.77ppm(d，3H)；0.73-0.55?ppm(m，2H)。
Step II I
At 3-[(1,4-Dioxa-spiral shell [4.5] last of the ten Heavenly stems-8-yl)-(anti-4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (401mg, 0.806mmol) the solution of tetrahydrofuran (THF) (4ml) in add 3N HCl solution (4ml), stirring at room reactant 20 hours.Use ethyl acetate (10ml) dilution then, separate organic layer, water ethyl acetate (2 * 10mL) washed twice.The ethyl acetate layer that merges is used Na with salt solution (10ml) washing
2SO
4Drying is filtered and is concentrated.Resistates obtains 315mg (86%) 3-[(4-methyl-cyclohexyl alkyl carbonyl by flash chromatography purifying (0-40%EtOAc/ hexane))-(4-oxygen-cyclohexyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters.
1H?NMR(CDCl
3，400MHz)7.64-7.62?ppm(m，2H)；7.48-7.40?ppm(m，3H)；7.02ppm(s，1H)；5.13-5.05?ppm(m，1H)；3.86?ppm(s，3H)；2.59-2.24?ppm(m，5H)；2.15-2.09?ppm(m，1H)；2.04-1.99?ppm(m，1H)；1.78-1.60?ppm(m，6H)；1.50-1.32ppm(m，3H)；0.78?ppm(d，3H)；0.74-0.57?ppm(m，2H)。
Step IV
With 3-[(4-methyl-cyclohexyl alkyl carbonyl)-(4-oxygen-cyclohexyl)-amino]-(34mg 0.075mmol) is dissolved in 4: 1 De diox: H to 5-phenyl-thiophene-2-carboxylic acid methyl esters
2In the O mixture (1ml), add then LiOH1N (375ul, 0.375mmol).After the stirring at room 3 hours, remove and desolvate, be distributed in the H that 5ml is acidified to pH 4 then
2Between O and the 5mlEtOAc.Separate organic layer, water ethyl acetate (2 * 5mL) washed twice.With the ethyl acetate layer drying (Na that merges
2SO
4) and concentrate.Resistates is by preparation type chromatography purification (10%MeOH/CH
2Cl
2), the 3-[(4-methyl-cyclohexyl alkyl carbonyl of acquisition 17mg (52%))-(4-oxygen-cyclohexyl)-amino]-5-phenyl-thiophene-2-carboxylic acid (compound 13).
1H?NMR(CD
3OD，400MHz)7.64-7.62?ppm(m，2H)；7.38-7.28?ppm(m，3H)；7.26ppm(s，1H)；4.88-4.81?ppm(m，1H)；2.55-2.41?ppm(m，1H)；2.26-1.91?ppm(m，3H)；1.88-1.26?ppm(m，11H)；0.88-0.78?ppm(m，1H)；0.69?ppm(d，3H)；0.63-0.48ppm(m，2H)。
Embodiment 15
3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid compound 15.
Step I
With 3-[(4-methyl-cyclohexyl alkyl carbonyl)-(4-oxygen-cyclohexyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (55mg 0.121mmol) is dissolved in the methyl alcohol (1.2ml), is cooled to 0 ℃, add then sodium borohydride (4.6mg, 0.121mmol).0 ℃ was stirred after 30 minutes, and with 10% hydrochloric acid soln (5ml) this reaction of going out suddenly, (3 * 10mL) extract water with ethyl acetate.With the ethyl acetate layer drying (Na that merges
2SO
4) and concentrate.Resistates is by preparation type chromatography purification (3%MeOH/CH
2Cl
2), the 3-[(that obtains 34mg (62%) is trans-4-hydroxyl-cyclohexyl)-(trans-4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters.
1H?NMR(CDCl
3，400MHz)：7.66-7.63?ppm(m，2H)；7.48-7.39?ppm(m，3H)；7.02ppm(s，1H)；4.62-4.54?ppm(m，1H)；3.85?ppm(s，3H)；3.46-3.39?ppm(m，1H)；2.03-1.91?ppm(m，4H)；1.83-1.78ppm(m，1H)；1.72-1.23ppm(m，10H)；1.07-0.97ppm(m，1H)；0.76?ppm(d，3H)；0.73-0.55?ppm(m，2H)。
Step II
With 3-[(trans-4-hydroxyl-cyclohexyl)-(trans-4-methyl-cyclohexyl alkyl carbonyl)-amino]-(34mg 0.075mmol) is dissolved in 4: 1 De diox: H to 5-phenyl-thiophene-2-carboxylic acid methyl esters
2In the O mixture (1ml), add then LiOH 1N (375ul, 0.375mmol).After the stirring at room 4 hours, remove and desolvate, be distributed in the H that 5ml is acidified to pH 4 then
2Between O and the 5mlEtOAc.Separate organic layer, water ethyl acetate (2 * 5mL) washed twice.The ethyl acetate layer drying (Na2SO4) that merges is also concentrated.Resistates is by preparation type chromatography purification (10%MeOH/CH
2Cl
2), the 3-[(that obtains 21mg (64%) is trans-4-hydroxyl-cyclohexyl)-(trans-4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid (compound 15).
1H?NMR(CD
3OD，400MHz)7.73-7.70?ppm(m，2H)；7.46-7.37?ppm(m，3H)；7.28ppm(s，1H)；4.47-4.41?ppm(m，1H)；3.38-3.32?ppm(m，1H)；2.14-2.08?ppm(m，1H)；1.99-1.88?ppm(m，4H)；1.80-1.77?ppm(bd，1H)；1.70-1.51?ppm(m，4H)；1,44-1.27ppm(m，5H)；1.10-1.03?ppm(m，1H)；0.77?ppm(d，3H)；0.72-0.55?ppm(m，2H)。
Embodiment 16
3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid compound 47.
Step I
With 3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (62mg, 0.136mmol) be dissolved in the benzene (0.7ml), add then p-nitrobenzoic acid (27mg, 0.163mmol) and triphenylphosphine (43mg, 0.163mmol).The solution that cooling forms in the ice bath, and the adding diethylazodicarboxylate (26 μ l, 0.163mmol).Stirring at room 22 hours; remove and desolvate; resistates obtains 3-{ (4-methyl-cyclohexyl alkyl carbonyl)-[4-(4-nitro-benzoyl oxygen)-cyclohexyl]-amino of 44mg (54%) by preparation type chromatography purification (30%EtOAc/ hexane) }-5-phenyl-thiophene-2-carboxylic acid methyl esters.
1H?NMR(CDCl
3，400MHz)7.91-7.83?ppm(m，4H)；7.69-7.64?ppm(m，2H)；7.50-7.47ppm(m，3H)；7.16?ppm(s，1H)；5.24?ppm(bs，1H)；4.82-4.74?ppm(m，1H)；3.86ppm(s，3H)；2.13-1.90?ppm(m，4H)；1.82-1.59?ppm(m，9H)；1.50-1.39?ppm(m，1H)；1.37-1.24?ppm(m，2H)；0.78?ppm(d，3H)；0.75-0.59?ppm(m，2H)。
Step II
With 3-{ (4-methyl-cyclohexyl alkyl carbonyl)-[4-(4-nitro-benzoyloxy)-cyclohexyl]-amino }-(44mg 0.073mmol) is dissolved in 4: 1 De diox: H to 5-phenyl-thiophene-2-carboxylic acid methyl esters
2Add then in the O mixture (1ml) LiOH 1N (365 μ l, 0.365mmol).After the stirring at room 4 hours, remove and desolvate, be distributed in the H that 5ml is acidified to pH 4 then
2Between O and the 5mlEtOAc.Separate organic layer, water ethyl acetate (2 * 5mL) washed twice.The ethyl acetate layer drying (Na2SO4) that merges is also concentrated.Resistates obtains 3-[(4-hydroxyl-cyclohexyl of 15mg (47%) by preparation type chromatography purification (10%MeOH/CH2Cl2))-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid.
1H?NMR(CD
3OD，400MHz)7.64-7.61?ppm(m，2H)；7.37-7.27?ppm(m，3H)；7.20ppm(s，1H)；4.43-4.37?ppm(m，1H)；3.79?ppm(bs，1H)；2.08-2.02?ppm(m，1H)；1.77-1,43?ppm(m，13H)；1.36-1.24?ppm(m，2H)；0.68?ppm(d，3H)；0.64-0.50ppm(m，2H)。
Embodiment 17
3-[(4-methoxyl group-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid compound 46.
Step I
With 3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (27mg, 0.059mmol) be dissolved among the THF (0.6ml), be cooled to 0 ℃ in the ice bath, add 60% sodium hydride (5mg, 0.118mmol), add the tetrabutylammonium iodide of catalytic amount subsequently.Stir after 1 hour, and the adding methyl iodide (37 μ l, 0.590mmol), reactant restir 3 hours.Water (5ml) goes out suddenly then, with ethyl acetate (3 * 5ml) extractions.With the ethyl acetate layer drying (Na that merges
2SO
4) and concentrate.Resistates is by preparation type chromatography purification (10%MeOH/CH
2Cl
2), 3-[(4-methoxyl group-cyclohexyl of acquisition 5mg (18%))-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid (compound 46).
1H?NMR(CDCl
3，400MHz)7.67-7.65?ppm(m，2H)；7.47-7.40?ppm(m，3H)；7.05ppm(s，1H)；4.59?ppm(bs，1H)；3.28?ppm(s，3H)；3.06-2.97?ppm(m，1H)；2.18-2.01ppm(m，4H)；1.94-1.90?ppm(m，1H)；1.74-1.25?ppm(m，11H)；0.77?ppm(d，3H)；0.71-0.61?ppm(m，2H)。
Embodiment 18
3-[(4-oxyimino-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid compound 16.
Step I
With 3-[(4-methyl-cyclohexyl alkyl carbonyl)-(4-oxygen-cyclohexyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (67mg 0.148mmol) is dissolved in the methyl alcohol (1.5ml), and the adding hydroxylamine hydrochloride (62mg, 0.888mmol).Stirring at room 2 hours reflux to stir 2 hours, added 10% sodium hydroxide solution, with the pH regulator of solution to 8-9.The solution that backflow forms 30 minutes is cooled to room temperature.Water (5ml) goes out suddenly then, with ethyl acetate (3 * 5ml) extractions.The ethyl acetate layer salt water washing that merges, dry (Na
2SO
4) and concentrate.Resistates obtains 3-[(4-oxyimino-cyclohexyl of 49mg (71%) by flash chromatography purifying (0-60%EtOAc/Hex))-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters.
1H?NMR(CDCl
3，400MHz)7.63-7.60?ppm(m，2H)；7.47-7.39?ppm(m，3H)；6,98ppm(s，1H)；4.90-4.82?ppm(m，1H)；3.84?ppm(s，3H)；3.39-3.29?ppm(m，1H)；2.44-2.20?ppm(m，2H)；2.13-2.09?ppm(m，1H)；2.04-1.73?ppm(m，4H)；1.70-1.57ppm(m，4H)；1.50-1.22?ppm(m，4H)；1.13-1.02?ppm(m，1H)；0.77?ppm(d，3H)；0.73-0.55?ppm(m，2H)。
Step II
With 3-[(4-oxyimino-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-(34mg 0.073mmol) is dissolved in 4: 1 De diox: H to 5-phenyl-thiophene-2-carboxylic acid methyl esters
2In O (1ml) mixture, add then LiOH 1N (365 μ l, 0.365mmol).After the stirring at room 3 hours, remove and desolvate, be distributed in the H that 5ml is acidified to pH 4 then
2Between O and the 5mlEtOAc.Separate organic layer, water ethyl acetate (2 * 5mL) washed twice.With the ethyl acetate layer drying (Na that merges
2SO
4) and concentrate.Resistates is by preparation type chromatography purification (10%MeOH/CH
2Cl
2), 3-[(4-oxyimino-cyclohexyl of acquisition 15mg (45%))-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid compound 16.
1H?NMR(CD
3OD，400MHz)7.72-7.69?ppm(m，2H)；7.50-7.36?ppm(m，3H)；7.29ppm(s，1H)；4.73-4.70?ppm(m，1H)；3.42-3.31?ppm(m，1H)；2.42-2.07?ppm(m，5H)；1.89-1.28?ppm(m，9H)；1.18-1.03?ppm(m，1H)；0.78?ppm(d，3H)；0.73-0.56ppm(m，2H)。
Prepare compound 25 in a similar way.
Embodiment 19
3-[(1-ethyl-3-methylamino--propyl group)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid compound 41.
Step I
3-amino-5-phenyl-thiophene-2-carboxylic acid methyl esters (1.0g, add in THF 4.29mmol) (1.0ml) stirred solution ketone (1.0g, 4.29mmol), dichloride dibutyl tin (130mg, 0.43mmol) and phenyl silane (582ul, 4.72mmol), stirring at room reaction mixture 2 days.Use saturated NaHCO then
3Solution this reaction of going out suddenly, mixture is with EtOAc extraction three times.Na is used in the extraction liquid salt water washing that merges
2SO
4Dry.Filter and concentrate.Crude product obtains 4-(2-methoxycarbonyl-5-phenyl-benzene sulphur-3-base the is amino)-piperidines-1-carboxylic acid benzyl ester of 1.86g (96%) by flash chromatography purifying (0-30%EtOAc/Hex).
Step II
(637mg, (2M is dissolved in CH to add oxalyl chloride in methylene dichloride 4.48mmol) (22ml) stirred solution in trans-4-methyl cyclohexane acid
2Cl
2, 4.5ml) solution adds 2 DMF subsequently.Stirring at room reaction mixture 2 hours, evaporation removes and desolvates and excessive oxalul chloride then.Crude product need not be further purified and be used for next step.
4-(2-methoxycarbonyl-5-phenyl-benzene sulphur-3-base amino)-piperidines-1-carboxylic acid benzyl ester (1.01g, add in ethylene dichloride 2.24mmol) (7.5ml) stirred solution trans-4-methylcyclohexyl chlorine (720mg, 4.48mmol).90 ℃ of 17 hours postcooling of reaction mixture that add thermosetting are used saturated NaHCO to room temperature
3Solution goes out suddenly, then with EtOAc extraction three times.Na is used in the extraction liquid salt water washing that merges
2SO
4Dry.Filter and concentrate.Crude product obtains 4-[(2-methoxycarbonyl-5-phenyl-benzene sulphur-3-yl of 1.00g (78%) by flash chromatography purifying (0-25%EtOAc/Hex))-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-piperidines-1-carboxylic acid benzyl ester.
1HNMR(CDCl
3，400MHz)7.65-7.62ppm(m，2H)；7.49-7.40ppm(m，3H)；7.31-7.23ppm(m，5H)；7.0?ppm(s，1H)；5.05?ppm(s，2H)；4.82-4.76?ppm(m，1H)；4.19ppm(bs，2H)；3.85?ppm(s，3H)；2.87?ppm(bs，2H)；2.03-1.58?ppm(m，9H)；1,49-1.28ppm(m，2H)；1.10?ppm(bs，2H)；0.78?ppm(d，3H)；0.73-0.56?ppm(m，2H)。
Step II I
At 4-[(2-methoxycarbonyl-5-phenyl-benzene sulphur-3-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-piperidines-1-carboxylic acid benzyl ester (506mg, 0.88mmol) 6: 1 ethyl acetate and methyl alcohol (7ml) mixture in add in the solution palladium on 10% activated carbon (103mg, 0.097mmol).The reaction mixture that forms is placed H
2Atmosphere (25psi), stirring at room 3 days is filtered on diatomite then, is evaporated to dried.Crude product is by flash chromatography purifying (100/90/16/1 CH
2Cl
2/ CHCl
3/ MeOH/Et
3N), obtain the 3-[(4-methyl-cyclohexyl alkyl carbonyl of 287mg (74%))-piperidin-4-yl-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters.
1H?NMR(CDCl
3，400MHz)7.64-7.61?ppm(m，2H)；7.46-7.38?ppm(m，3H)；7.04ppm(s，1H)；4.74-4.68?ppm(m，1H)；3.84?ppm(s，3H)；3.13-3.03?ppm(m，2H)；2.78-2.67?ppm(m，2H)；2.03-1.92?ppm(m，2H)；1.77-1.74?ppm(m，1H)；1.69-1.13ppm(m，9H)；0.77?ppm(d，3H)；0.72-0.59?ppm(m，2H)。
Step IV
With 3-[(4-methyl-cyclohexyl alkyl carbonyl)-piperidin-4-yl-amino]-(20mg 0.045mmol) is dissolved in 4: 1 De diox: H to 5-phenyl-thiophene-2-carboxylic acid methyl esters
2In the O mixture (0.5ml), add then LiOH1N (135 μ l, 0.135mmol).Stirring at room 4 hours, is removed then and is desolvated to pH 3-4 with 10% hydrochloric acid soln acidified reaction mixture.Further use cold water (1ml) dilution, and filter, obtain the 3-[(4-methyl-cyclohexyl alkyl carbonyl of 16mg (84%))-piperidin-4-yl-amino]-5-phenyl-thiophene-2-carboxylic acid (compound 41).
1HNMR(CD
3OD，400MHz)7.78-7.75?ppm(m，2H)；7.50-7.41?ppm(m，4H)；4.77-4.69ppm(m，1H)；3.47-3.36?ppm(m，2H)；3.16-3.06?ppm(m，2H)；2.24-2.21?ppm(m，1H)；2.15-2.07?ppm(m，2H)；1.91-1.80?ppm(m，1H)；1.76-1.51?ppm(m，6H)；1,44-1.26ppm(m，2H)；0.79?ppm(d，3H)；0.76-0.60?ppm(m，2H)。
Embodiment 20
3-[(1-(benzyl-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino)-5-phenyl-thiophene-2-carboxylic acid compound 52.
Step I
At 3-[(4-methyl-cyclohexyl alkyl carbonyl)-piperidin-4-yl-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (43mg, 0.098mmol) ethylene dichloride (1.0ml) solution in add phenyl aldehyde (15 μ l, 0.146mmol), add subsequently sodium triacetoxy borohydride (41mg, 0.195mmol).Stirring at room reaction mixture 4 hours is used saturated NaHCO
3Solution goes out suddenly, uses ethyl acetate (3 * 5ml) extractions then.The extract salt water washing that merges, and use Na
2SO
4Dry.Filter and concentrate.Resistates obtains 3-[(1-benzyl-piperidin-4-yl of 32mg (61%) by preparation type chromatography purification (50%EtOAc/Hex))-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters.
Step II
With 3-[(1-benzyl-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-(32mg 0.060mmol) is dissolved in 4: 1 De diox: H to 5-phenyl-thiophene-2-carboxylic acid methyl esters
2In the O mixture (0.5ml), add then LiOH 1N (180ul, 0.180mmol).Stirring at room 2 hours refluxes and stirs after 4 hours, removes and desolvates, and is distributed in the H that 5ml is acidified to pH 4 then
2Between O and the 5mlEtOAc.Separate organic layer, water ethyl acetate (2 * 5mL) washed twice.With the ethyl acetate layer drying (Na that merges
2SO
4) and concentrate.Resistates is by preparation type chromatography purification (10%MeOH/CH
2Cl
2), 3-[(1-(benzyl-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino of acquisition 22mg (71%))-5-phenyl-thiophene-2-carboxylic acid (compound 52).
1H?NMR(CD
2Cl
2，400MHz)7.41-7.36?ppm(m，4H)；7.31-7.28?ppm(m，6H)；6,82ppm(s，1H)；4.80-4.73?ppm(m，1H)；4.37?ppm(d，1H)；3.65?ppm(bd，1H)；3.53ppm(d，1H)；3.10?ppm(bd，1H)；2.63?ppm(t，1H)；2.47?ppm(t，1H)；2.10-2.06ppm(m，2H)；1.85-1.63?ppm(m，4H)；1.57-1.38?ppm(m，4H)；1.28-1.18?ppm(m，2H)；0.66?ppm(d，3H)；0.62-0.50?ppm(m，2H)。
Embodiment 21
3-[(1-ethanoyl-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid compound 48.
Step I
At 3-[(4-methyl-cyclohexyl alkyl carbonyl)-piperidin-4-yl-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (58mg, 0.132mmol) methylene dichloride (1.3ml) solution in add pyridine (64ul, 0.789mmol), add subsequently diacetyl oxide (50ul, 0.526mmol) and the DMAP of catalytic amount.The solution that stirring at room forms 18 hours is used NaHCO then
3Saturated solution (5ml) goes out suddenly.Water phase separated is with ethyl acetate (2 * 5ml) washings.With the organic layer drying (Na that merges
2SO
4), filter and concentrate.Resistates is by preparation type chromatography purification (100/90/16/1CH
2Cl
2/ CHCl
3/ MeOH/Et
3N), obtain 50mg (78%) 3-[(1-ethanoyl-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters.
Step II
With 3-[(1-ethanoyl-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-(50mg 0.104mmol) is dissolved in 4: 1 De diox: H to 5-phenyl-thiophene-2-carboxylic acid methyl esters
2In the O mixture (1ml), add then LiOH 1N (310ul, 0.310mmol).Stirring at room 5 hours, and reflux and stirred 4 hours, remove and desolvate, be distributed in the H that 5ml is acidified to pH 4 then
2Between O and the 5mlEtOAc.Separate organic layer, water ethyl acetate (2 * 5mL) washed twice.With the ethyl acetate layer drying (Na that merges
2SO
4) and concentrate.Resistates is by preparation type chromatography purification (100/90/16/1 CH
2Cl
2/ CHCl
3/ MeOH/Et
3N), obtain 3-[(1-ethanoyl-piperidin-4-yl of 27mg (56%))-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid (compound 48).
1H NMR (CD
3OD, 400MHz) 1: 1 rotational isomer mixture 7.75-7.73 ppm (m, 2H); 7.48-7.40 ppm (m, 3H); 7.37 ppm (s, 1H); 4.74-4.51 ppm (m, 1H); 3.99-3.90ppm (m, 1H); 3.23-3.12 ppm (m, 1H); 2.70-2.60 ppm (m, 1H); 2.27-2.00 ppm (m, 2H); 2.04 ppm (s, 1.5H); 2.00 ppm (s, 1.5H); 1.96-1.87 ppm (m, 1H); 1.77-1.06ppm (m, 9H); 0.78 ppm (d, 3H); 0.73-0.57 ppm (m, 2H).
Prepare compound 63 in a similar way.
1H NMR (CD
3OD, 400MHz) 1: 1 rotational isomer mixture 7.91 ppm (s, 0.5H); 7.88ppm (s, 0.5H); 7.75-7.72 ppm (m, 2H); 7.48-7.39 ppm (m, 3H); 7.37 ppm (s, 0.5H); 7.36 ppm (s, 0.5H); 4.78-4.68 ppm (m, 1H); 4.42-4.31 ppm (m, 1H); 3.79-3.63ppm (m, 1H); 3.26-3.15 ppm (m, 1H); 2.78-2.66 ppm (m, 1H); 2.12-1.91 ppm (m, 3H); 1.76-1.04 ppm (m, 9H); 0.78 ppm (d, 3H); 0.73-0.57 ppm (m, 2H).
As preparation compound 23 as described in the embodiment 24, compound 39, compound 40.
Compound 23:
1H NMR (CD
3OD, 400MHz) 7.65-7.63 ppm (m, 2H); 7.25 ppm (s, 1H); 7.16-7.12 ppm (m, 2H); 4.76-4.70 ppm (m, 1H); 3.53-3.46 ppm (m, 1H); 3.17-3.04ppm (m, 2H); 2.80 ppm (s, 3H); 2.23-2.09 ppm (m, 3H); 2.01-1.92 ppm (m, 1H); 1.82-1.79 ppm (m, 1H); 1.70-1,48 ppm (m, 5H); 1, and 41-1.25 ppm (m, 2H); 0.77ppm (d, 3H); 0.73-0.54 ppm (m, 2H).
Compound 39:
1HNMR (CD
3OD, 400MHz) 7.77-7.74ppm (m, 2H); 7.50-7.40 ppm (m, 4H); 4.77-4.71 ppm (m, 1H); 3.62-3.54 ppm (m, 2H); 3.15-3.04 ppm (m, 4H); 2.29-2.08ppm (s, 3H); 1.97-1.88 ppm (m, 1H); 1.78-1.51 ppm (m, 6H); 1, and 45-1.34 ppm (m, 1H); 1.30-1.26 ppm (m, 1H); 1.28 ppm (t, 3H); 0.79 ppm (d, 3H); 0.76-0.59 ppm (m, 2H).
Compound 40:
1H NMR (CD
3OD, 400MHz) 7.76-7.74 ppm (m, 2H); 7.49-7.40ppm (m, 4H); 4.78-4.72 ppm (m, 1H); 3.51-3.42 ppm (m, 3H); 3.22-3.13 ppm (m, 2H); 2.30-2.09 ppm (s, 3H); 2.00-1.91 ppm (m, 1H); 1.78-1.51 ppm (m, 7H); 1, and 47-1.34ppm (m, 1H); 1.30 ppm (d, 6H); 0.79 ppm (d, 3H); 0.74-0.59 ppm (m, 2H).
Embodiment 22
3-[(4-methyl-cyclohexyl alkyl carbonyl)-(1-methyl isophthalic acid-oxygen-piperidin-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid compound 65.
Step I
With 3-[(4-methyl-cyclohexyl alkyl carbonyl)-(1-methyl-piperidin-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (42mg 0.092mmol) is dissolved in the methylene dichloride (1.8ml), add then m-chlorine peroxybenzoic acid (27mg, 0.111mmol).After the stirring at room 2 hours, remove and desolvate, resistates dilutes with ethyl acetate (5ml).This solution is used 10% sodium hydroxide solution again, and (2 * 5ml), salt solution (5ml) washs, dry then (Na
2SO
4), filter and concentrate.Resistates is by preparation type chromatography purification (10%MeOH/CH
2Cl
2), the 3-[(4-methyl-cyclohexyl alkyl carbonyl of acquisition 33mg (77%))-(1-methyl isophthalic acid-oxygen-piperidin-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters.
1H?NMR(CDCl
3，400MHz)7.64-7.61?ppm(m，2H)；7.45-7.37?ppm(m，3H)；7.10ppm(s，1H)；4.76-4.68?ppm(m，1H)；3.83?ppm(s，3H)；3.34-3.22?ppm(m，4H)；3.18ppm(s，3H)；2.85?ppm(bs，2H)；2.57-2.47?ppm(m，1H)；2.23-2.14?ppm(m，1H)；2.03-1.96?ppm(m，1H)；1.90-1.87?ppm(m，1H)；1.70-1.58?ppm(m，3H)；1,46-1.35ppm(m，1H)；1.31-1.22?ppm(m，2H)；0.76?ppm(d，3H)；0.71-0.56?ppm(m，2H)。
Step II
With 3-[(4-methyl-cyclohexyl alkyl carbonyl)-(1-methyl isophthalic acid-oxygen-piperidin-4-yl)-amino]-(33mg 0.070mmol) is dissolved in 4: 1 De diox: H to 5-phenyl-thiophene-2-carboxylic acid methyl esters
2In the O mixture (0.7ml), add then LiOH 1N (210 μ l, 0.210mmol).After the stirring at room 4 hours, to pH 3-4, remove then and desolvate with 10% hydrochloric acid soln acidified reaction mixture.With further dilution of cold water (1ml), obtain the 3-[(4-methyl-cyclohexyl alkyl carbonyl of 23mg (72%) after the filtration)-(1-methyl isophthalic acid-oxygen-piperidin-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid (compound 65).
1H?NMR(CD
3OD，400MHz)7.74-7.72?ppm(m，2H)；7.48-7.39?ppm(m，3H)；7.38ppm(s，1H)；4.75-4.67?ppm(m，1H)；3.80-3.65?ppm(m，4H)；3.42?ppm(s，3H)；2.32-2.21?ppm(m，1H)；2.17-1.95?ppm(m，4H)；1.81-1.71?ppm(m，2H)；1.66-1.28ppm(m，5H)；0.78?ppm(d，3H)；0.76-0.59?ppm(m，2H)。
Embodiment 23
3-[(2-hydroxy-4-methyl-hexanaphthene carbonyl)-(tetrahydrochysene-pyrans-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid compound 31.
Step I
As embodiment 25, step VI is described, and with being dissolved in 1, the triphenylphosphine of 2-ethylene dichloride carries out acylations under refluxing.
Step II
With 3-[(2-hydroxy-4-methyl-hexanaphthene carbonyl)-(tetrahydrochysene-pyrans-4-yl)-amino]-(31mg 0.062mmol) is dissolved in 4: 1 De diox: H to 5-phenyl-thiophene-2-carboxylic acid methyl esters
2In the O mixture (0.6ml), add then LiOH 1N (310ul, 0.310mmol).After the stirring at room 4 hours, remove and desolvate, be distributed in the H that 5ml is acidified to pH 4 then
2Between O and the 5mlEtOAc.Separate organic layer, water ethyl acetate (2 * 5mL) washed twice.With the ethyl acetate layer drying (Na that merges
2SO
4) and concentrate.Resistates obtains the 3-[(2-hydroxy-4-methyl-5 hexanaphthene carbonyl of 14mg (52%) by the preparation HPLC purifying)-(tetrahydrochysene-pyrans-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid (compound 31).
1H?NMR(CD30D，400MHz)7.80-7.75?ppm(m，2H)；7.50-7.40?ppm(m，4H)；4.80-4.68?ppm(m，1H)；4.15?ppm(s，1H)；4.00-3.85?ppm(m，2H)；3.55-3.40ppm(m，2H)；2.35-2.15?ppm(m，1H)；2.00-1,45?ppm(m，4H)；1,40-1.25?ppm(m，2H)；0.75?ppm(d，3H)；0.73-0.55?ppm-(m，2H)。
Embodiment 24
Chlorination 4-[(2-carboxyl-5-phenyl-benzene sulphur-3-yl)-(trans-4-methyl-cyclohexyl alkyl carbonyl)-amino]-1-methyl-piperidine compounds 11.
Step I
(a) in 30 minutes, 1-methyl-piperidin-4-one-(6.0g, 53mmol, 6.52mL) and Et
3N (14.16g, 140mmol, 19.5mL) 1, drip in 4-diox (20mL) stirred solution chlorine trimethyl silane (7.6g, 70mmol, 8.88mL).Slowly add reaction mixture to the 110 ℃ backflow of thermosetting, under same temperature, stirred 24 hours, add chlorine trimethyl silane (4.44mL) again, heat and (got its five equilibrium and carry out in 24 hours
1HNMR), be cooled to room temperature, filter out solid.Wash with Skellysolve A.Filtrate concentrates in rotatory evaporator, then with Skellysolve A dilution, solids removed by filtration.On rotatory evaporator,, provide 1-methyl-4-trimethylsiloxy-1,2,3 with the solution that produces with after high vacuum concentrates, 6-tetrahydrochysene-pyridine (9.68g,
1HNMR shows the silicomethane alcohol ether (silylenolether) and the initial substance of about 10: 1 ratios).
Crude product needn't be further purified and can be used for next step.
(b) at methyl-3-amino-5-phenyl thiophene-carboxylicesters (233mg, 1.0mmol) and 1-methyl-4-trimethylsiloxy-1,2,3, in the stirred solution of the ethylene dichloride (3.0mL) of 6-tetrahydrochysene-pyridine (370mg.2.0cymol), adding AcOH (0.1mL, 2.0eq), a collection of subsequently adding NaBH (OAc)
3(424mg, 2.0mmol).The reaction mixture that forms in stirring at room adds the 10%NaOH aqueous solution (up to alkalescence) to weekend, after 30 minutes, uses the dichloromethane extraction reaction mixture.Organic extract liquid salt water washing after drying.Crude product carries out purifying, subsequently CHCl with the 20%EtOAc/ hexane to unreacted initial substance by silicagel column
3/ MeOH/Et
3N (180/16/1) purifying obtains 3-(1-methyl-piperidin-4-yl amino)-5-phenyl-thiophene-2-carboxylic acid methyl esters (240mg, 73%).NMR
1H(CDCl
3，400MHz)：7.64-7.6(m，2H)，7.43-7.34(m，3H)，6.83(brs，2H)，3.83(s，3H)，3.46-3.4(m，1H)，2.82-2.74(m，1H)，2.3(s，3H)，2.26-2.2(m，4H)，1.72-1.62(m，2H)。
Step II
(a) at trans-4-methyl cyclohexane acid (656mg, 4.6mmol) methylene dichloride (23mL) stirred solution in add oxalyl chloride (2M, 4.6mL) dichloromethane solution, add 2-3 subsequently and drip DMF (with 22 G pins), stir after 2 hours, remove on rotatory evaporator and desolvate and excessive oxalul chloride, rough vacuum is removed trace solvent, and (note: product is a high volatile volatile, can not use vacuum for a long time, about 1-2 minute).Thick 4-methyl-cyclohexyl alkane dicarbonyl chloride is used for next step at once.
(b) 3-(1-methyl-piperidin-4-yl amino)-5-phenyl-thiophene-2-carboxylic acid methyl esters (540mg, 1.636mmol) 1, in 2-ethylene dichloride (15mL) stirred solution, add trans-4-methyl-cyclohexyl alkane dicarbonyl chloride, add PPh subsequently
3(429mg, 1.635).The reaction mixture that forms is cooled to room temperature 90 ℃ of heating 48 hours, with the alkalization of the 10%NaOH aqueous solution, uses dichloromethane extraction then.With the organic extract liquid that merges salt water washing after drying, concentrate, purifying on silica gel column chromatography is with 200/90/16/1 (CH
2Cl
2/ CHCl
3/ MeOH/Et
3N) at first wash-out go out 3-[(trans-4-methyl-cyclohexyl alkyl carbonyl)-(1-methyl-piperidin-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (760mg, pollute have cyclohexylenedinitrilotetraacetic acid), follow by initial substance (270mg).NMR
1H(CDCl
3，400MHz)：7.64-7.6(m，2H)，7.47-7.38(m，3H)，7.04(s，1H)，4.68-4.58(m)，3.84(s，3H)，2.95-2.8(m，2H)，2.26(s，3H)，2.2-1.26(m，14H)，0.767(d，J＝6.6，3H)，0.74-0.56(m，2H)。
Step II I
With 3-[(trans-4-methyl-cyclohexyl alkyl carbonyl)-(1-methyl-piperidin-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (176mg, 0.387mmol) and LiOH monohydrate (48.8mg, 1.16mmol, 4.0 eq) Zai diox: water (3: 1.3,9mL, 0.1M) in 50 ℃ in mixture heating 5 hours, be cooled to room temperature, with the acidifying of 1N HCl solution, concentrate,, filter out product with the less water dilution, dry (136mg) then, with the hexane development for several times, to remove the acid of 4-methyl cyclohexane, obtain chlorination 4-[(2-carboxyl-5-phenyl-benzene sulphur-3-yl)-(trans-4-methyl-cyclohexyl alkyl carbonyl)-amino]-1-methyl-piperidines (compound 11), 101mg, 60% productive rate).
NMR
1H(CD
3OD，400MHz)：7.76-7.72(m，2H)，7.5-7.38(m，4H)，4.8-4.65(m，1H)，3.6-3.4(m，2H)，3.25-3.2(m，2H)，2.8(s，3H)，2.3-1.2(m，12H)，0.78(d，J＝6.6Hz，3H)，0.96-0.58(m，2H)。
Embodiment 25
(4R)-5-(4-fluoro-phenyl)-3-[sec.-propyl-(4-methyl-cyclohexyl-1-alkene carbonyl)-amino]-thiophene-2-carboxylic acid compound 26 and (1R, 2S, 4R)-5-(4-fluoro-phenyl)-3-[(2-hydroxy-4-methyl-hexanaphthene carbonyl)-sec.-propyl-amino]-thiophene-2-carboxylic acid compound 24.
Compound (1S, 2R, 5R)-2-sec.-propyl-5-methyl-cyclohexyl alcohol is according to Tetrahedron Letter, and (1993), vol.49 prepares described in the pp6429-6436.
NMR
1H(CDCl
3，400MHz)：4.95?ppm(s，1H)；4.78?ppm(s，1H)；3.99?ppm(s，1H)；2.01-1.95?ppm(m，2H)；1.79?ppm(s，3H)；1.79-1.66?ppm(m，3H)；1,48-1,42ppm(m，1H)；1.16-1.09?ppm(m，1H)；1.00-0.87?ppm(m，2H)；0.88?ppm(d，3H)。
Step I
At-78 ℃, (1S in being dissolved in DCM (67mL) and MeOH (1.6mL), 2R, 5R)-and 2-sec.-propyl-5-methyl-cyclohexyl alcohol (2.07g, 13.42mmol) bubbling adds ozone/oxygen gas, becomes blueness up to reaction mixture, fall excessive ozone with the oxygen purge, add methyl-sulfide (4.9mL) under same temperature, slowly be heated to room temperature, stirring is spent the night, concentrate, on column chromatography, use 10-20%EtOAc/ hexane purifying, obtain (1R, 2S, 4R)-and 1-(2-hydroxy-4-methyl-cyclohexyl)-ketene (1.40g, 67%), be oily matter.
NMR
1H(CDCl
3，400MHz)：4.29-4.27?ppm(m，1H)；2.40-2.35?ppm(m，1H)；2.19ppm(s，3H)；1.91-1.73?ppm(m，5H)；1.05-0.91?ppm(m，2H)；0.88?ppm(d，3H)。
Step II
NaOH (4.8, g, the 119.2mmol) aqueous solution (40mL) and 1, add in the ice-cooled solution of 4-diox (30mL) bromine (1.5mL, 29.57mmol).In the NaOBr yellow solution that produces, drip (1R, 2S, 4R)-1-(2-hydroxy-4-methyl-cyclohexyl)-ketene (1.4g, 8.962mmol) De diox (130mL) solution and water (35mL).The solution of 10-15 ℃ of stirring formation 3 hours.Add Na
2SO
3(1.1g is in the 11mL water) solution decomposing excessive NaOBr solution is used the 10%HCl acidifying, extracts with DCM.With the organic extract liquid salt water washing that merges, dry and concentrated, obtain (1R, 2S, 4R)-2-hydroxy-4-methyl-hexahydrobenzoic acid (1.30g, 92%).NMR
1H(CDCl
3，400MHz)：4.34?ppm(s，1H)；2.43-2.39?ppm(m，1H)；1.96-1.76?ppm(m，5H)；1.14-1.08?ppm(m，1H)；1.02-0.93?ppm(m，1H)；0.90ppm(d，3H)。
Step II I
(1R, 2S, 4R)-2-hydroxy-4-methyl-hexahydrobenzoic acid (162mg, add in methylene dichloride 1.02mmol) (5ml) solution pyridine (495 μ l, 6.12mmol), add subsequently diacetyl oxide (385 μ l, 4.08mmol).Stirring at room reaction mixture 20 hours.Remove then and desolvate, add 10ml 3N HCl solution.Stir this mixture 30 minutes, and slowly added NaHCO then
3Saturated solution is up to pH=9-10.This solution is used ethyl acetate (2 * 5ml) extractions then.Water is with the acidifying of 10%HCl solution, with ethyl acetate (3 * 5ml) extractions.The combined ethyl acetate layer, dry (Na
2SO
4), concentrate, acquisition 109mg (53%) (1R, 2S, 4R)-2-acetoxyl group-4-methyl-cyclohexyl alkane carboxylic acid.
NMR
1H(CDCl
3，400MHz)：4.34-4.32?ppm(m，1H)；2.42-2.37?ppm(m，1H)；1.95-1.76?ppm(m，5H)；1.13-1.06?ppm(m，1H)；1.01-0.92?ppm(m，1H)；0.89ppm(d，3H)。
Step IV
(1R, 2S, 4R)-(109mg, (545ul 1.09mmol), adds 1 dimethyl formamide to 2-acetoxyl group-4-methyl-cyclohexyl alkane carboxylic acid subsequently to add oxalyl chloride in methylene dichloride 0.54mmol) (2.7ml) solution.Stirring at room reaction mixture 4 hours.Remove then and desolvate, and acquisition 119mg (99%) chlorination (1R*, 2S*, 4R*)-2-acetoxyl group-4-methyl-cyclohexyl alkane carboxylic acid.NMR
1H(CDCl
3，400MHz)：5.45?ppm(s，1H)；2.46-2.42?ppm(m，1H)；2.02?ppm(s，3H)；2.02-1.96?ppm(m，1H)；1.91-1.76ppm(m，3H)；1.70-1.61?ppm(m，1H)；1.16-1.08?ppm(m，1H)；0.99-0.88?ppm(m，1H)；0.87?ppm(d，3H)。
Step V
At 3-amino-5-(4-fluoro-phenyl)-thiophene-2-carboxylic acid methyl esters (0.502g, 2.0mmol) 1, in 2-ethylene dichloride (6.0mL) stirred solution order add 2-methoxyl group propylene (0.38mL, 4.0mmol), AcOH (0.114mL, 2.0mmol) and NaBH (OAc)
3(0.84g 4.0mmol), stirred 2 hours.Use EtOAc and H then
2The O dilution.Add NaHCO
3, this aqueous solution is adjusted to pH=7.Water extracts with EtOAc, and MgSO is used in the salt water washing of the extraction liquid of merging
4Drying is filtered.With hexane to the 10%EtOAc-hexane in conjunction with the elutriant purifying, obtain 5-(4-fluoro-phenyl)-3-sec.-propyl aminothiophene-2-carboxylate methyl ester (0.538g, 92% productive rate).
Step VI:(4R)-5-(4-fluoro-phenyl)-3-[sec.-propyl-(4-methyl-cyclohexyl-1-alkene carbonyl)-amino]-thiophene-2-carboxylic acid methyl esters and (1R, 2S, 4R)-3-[(2-acetoxyl group-4-methyl-cyclohexyl alkyl carbonyl)-sec.-propyl-amino]-5-(4-fluoro-phenyl)-thiophene-2-carboxylic acid methyl esters
At 5-(4-fluoro-phenyl)-3-sec.-propyl amino-thiophene-2-carboxylic acid methyl esters (146mg, 0.50mmol) 1, add in 2-ethylene dichloride (1.5ml) solution and be dissolved in 1, chlorination (1R in the 2-ethylene dichloride (0.5ml), 2S, 4R)-(119mg 0.54mmol), adds PPh to 2-acetoxyl group-4-methyl-cyclohexyl alkane carboxylic acid subsequently
3(131mg, 0.5mmol).The solution of 90 ℃ of stirring formation 24 hours is cooled to room temperature then.With ethyl acetate (10ml) and NaHCO
3Saturated solution (10ml) dilution.Water phase separated, (2 * 10ml) washings are with the organic layer drying (Na that merges with ethyl acetate
2SO
4), filter and concentrate.Resistates obtains the mixture of 96mg title compound by flash chromatography purifying (0-25%EtOAc/ hexane).
Step VII
The compound (95mg) of step VI is dissolved in diox: H
2In the mixture (1.0mL) of O (4: 1), add 600 μ l LiOH 1N.60 ℃ after 24 hours, reaction mixture is cooled to room temperature, remove and desolvate.Resistates is distributed in the H that 10ml is acidified to pH 4
2Between O and the 10ml EtOAc.Separate organic layer, (2 * 10ml) wash water with ethyl acetate.With the ethyl acetate layer drying (Na that merges
2SO
4), concentrate, resistates obtains (4R)-5-(4-fluoro-phenyl)-3-[sec.-propyl-(4-methyl-cyclohexyl-1-alkene carbonyl)-amino by preparation type chromatography purification]-thiophene-2-carboxylic acid (compound 26) is (21mg).NMR
1H(CD
3OD，400MHz)：
7.76-7.68 (m, 2H), 7.3-7.1 (m, 3H), 5.78 (brs, 1H), 4.9-4.75 (m, 1H), 2.3-1,4 (m), 1.33 (d, J=4.3.3H), 1.09 (d, J=4.5.3H), 0.815 (d, J=3.5,3H) and (1R, 2S, 4R)-5-(4-fluoro-phenyl)-3-[(2-hydroxy-4-methyl-hexanaphthene carbonyl)-sec.-propyl-amino]-thiophene-2-carboxylic acid (compound 24) (41mg).NMR
1H(CD
3OD，400MHz)：7.75-7.7(m，2H)，7.23(s，1H)，7.2-7.15(m，2H)，4.9-4.8(m，1H)，2.0-1,4(m，5H)，1.206(d，J＝6.6，3H)，1.017(d，J＝6.4.3H)，0.76(d，J＝6.6，3H)。
Embodiment 26
3-[sec.-propyl-(5-methyl-3.6-dihydro-2H-pyrans-2-carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid compound 30.
Step I
NaH (55% is dispersed in the oil, 227.9mg, 5.2mmol, hydroxyl-amylene-(0.576g, THF 4.0mmol) (20mL) solution stirred 1 hour the 4-acetoacetic ester to drip 2-in the stirred suspension of THF 1.3eq) (20mL) cold (0 ℃).Reaction mixture is used 3-bromo-2-methacrylic then, and (0.81g, 6.0mmol 0.61mL) handle, and slowly are heated to room temperature, stir 1 hour.Use NH
4The Cl saturated solution carefully goes out suddenly.(3 * 20mL) extract reaction mixture, organic solution salt water washing, dry (Na with EtOAC
2SO
4), concentrate.Resistates with 5%EtOAc-hexane purifying, obtains 2-(2-methyl-allyloxy)-amylene-4-acetoacetic ester (0.521g, 66%) on silica gel column chromatography, be oily matter.NMR
1H(CDCl
3，400MHz)：5.9-5.78(m，1H)，5.16-5.06(m，2H)，4.97(s，1H)，4.91(s，1H)，4.26-4.16(m，2H)，4.072(d，J＝12.3，1H)，3.93(t，J＝6.5，1H)，3.813(d，J＝12.4，1H)，1.15(s，3H)，1.28(t，J＝7.2，3H)。
Step II
At 2-(2-methyl-allyloxy)-amylene-4-acetoacetic ester (396mg, CH 2.0mmol)
2Cl
2In (100mL, 0.02M solution) backflow stirred solution, (85mg is 0.1mmol) at CH for (benzylidene) ruthenium (IV) to drip tricyclohexyl phosphine (dichloride 1,3-two (2,4, the 6-trimethylphenyl)-4,5-glyoxalidine-2-yl)
2Cl
2Solution (3.0mL).After 50 minutes, reaction mixture is cooled to room temperature, concentrates, in conjunction with the elutriant purifying, use EtOAc/ hexane (1: 20) in conjunction with the wash-out purifying, obtains 5-methyl-3 as elutriant at silicagel column, 6-dihydro-2H-pyrans-2-carboxylic acid, ethyl ester (320mg, 92% productive rate) is brown oil.NMR
1H(CDCl
3，400MHz)：5.51(br?s，1H)，4.28-4.08(m，4H)，2.32(brs，3H)，1.29(t，J＝7.2，3H)。
Step II I
With 5-methyl-3,6-dihydro-2H-pyrans-2-carboxylic acid, ethyl ester (140mg, MeOH 0.823mmol) (3.5mL) and the 10%NaOH aqueous solution (1.0mL, 2.5mmol) in solution 65 ℃ the heating 3 hours, cool off this reaction mixture to room temperature, solvent evaporated, dilute with water.The aqueous solution washs with ether, with the acidifying of 1N HCl solution, extracts with ether.Ethereal solution is also dry with the salt water washing.Solvent evaporated obtains 5-methyl-3,6-dihydro-2H-pyrans-2-carboxylic acid (82mg, 70% productive rate).
NMR
1H(CDCl
3，400MHz)：5.54-5.5(m，1H)，4.24-4.1(m，3H)，2,4-2.3(m，2H)，1.6(s，3H)。
Step IV
Use PPh
3(78.6mg, 0.3mmol) and NCS (39.9mg, 0.3mmol), coupling 3-sec.-propyl amino-5-phenyl-thiophene-2-carboxylic acid methyl esters (63.3mg, 0.23mmol) and 5-methyl-3, and 6-dihydro-2H-pyrans-2-carboxylic acid (40mg, 0.28mmol), obtain 3-[sec.-propyl-(5-methyl-3.6-dihydro-2H-pyrans-2-carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (60mg, 65.3% productive rate).
NMR
1H (CDCl
3, 400MHz is for main rotational isomer): 7.68-7.62 (m, 2H), 7.4-7.5 (m, 3H), 7.22 (s, 1H), 5.46 (m, 1H), 3.86 (s, 3H), 1,48 (s, 3H), 1.24 (d, 3H), 1.0 (d, 3H)
Step V
As described in embodiment 25 steps 7, use LiOHH
2O (12mg) hydrolysis 3-[sec.-propyl-(5-methyl-3.6-dihydro-2H-pyrans 2-carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (38mg, 0.095mmol), obtain 3-[sec.-propyl-(5-methyl-3.6-dihydro-2H-pyrans-2-carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid (compound 30) (13mg, 35.5% productive rate).NMR
1H (CD
3OD, 400MHz is for main rotational isomer): 7.8-7.7 (m, 2H), 7.5-7.3 (m, 4H), 5.45 (brs, 1H), 4.95-4.8 (m, 1H), 2.56-1.82 (m), 1,46 (brs, 3H), 1.26 (d, 3H), 1.2 (d, 3H), 1.0-0.84 (m).
Embodiment 27
3-[sec.-propyl-(cis-5-methyl-tetrahydrochysene-pyrans-2-carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid compound 29 and 3-[sec.-propyl-(trans-5-methyl-tetrahydrochysene-pyrans-the 2-carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid compound 27.
Step I
At 5-methyl-3, (40mg adds 5%Pt-C (20mg) in MeOH 0.28mmol) (2.0mL) solution, 20psi hydrogenation 16 hours to 6-dihydro-2H-pyrans-2-carboxylic acid.Go out reaction mixture by diatomite filtration,, obtain the geometrical isomer (37mg, 91%) of 5-methyl-tetrahydrochysene-pyrans-2-carboxylic acid of 2: 1 with MeOH washing, concentrated filtrate.
Step II
Adopt embodiment 26, the described method of step 4 obtains separable 3-[sec.-propyl-(cis-5-methyl-tetrahydrochysene-pyrans-2-carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (30mg, 35.5%) mixture.NMR
1H (CDCl
3, 400MHz is for main rotational isomer): 7.66-7.6 (m, 2H), 7.48-7.36 (m, 3H), 7.162 (s, 1H), 5.0-4.88 (m, 1H), 3.98-3.94 (m), 3.86 (s, 3H), 3.29 (m), 2.18-1,4 (m), 1.25 (d, 3H), 0.98 (d, 3H), 0.72 (d is 3H) with 3-[sec.-propyl-(trans-5-methyl-tetrahydrochysene-pyrans-the 2-carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (14.0mg, 16.6%).NMR
1H (CDCl
3, 400MHz is for main rotational isomer): 7.65-7.63 (m, 2H), 7.48-7.38 (m, 3H), 7.184 (s, 1H), 4.96-4.86 (m, 1H), 3.86-3.52 (m), 2.55 (t, 1H), 1.96-1,46 (m), 1.218 (d, J=3.7.3H), 0.985 (d, J=6.7.3H), 0.657 (d, J=6.7.3H).
Step II I
As described in embodiment 25 steps 7, use LiOHH
2O hydrolysis 3-[sec.-propyl-(cis-5-methyl-tetrahydrochysene-pyrans-2-carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (30mg, 0.075mmol), obtain 3-[sec.-propyl-(cis-5-methyl-tetrahydrochysene-pyrans-2-carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid (compound 29) (13mg, 44.8%).NMR
1H (CD
3OD, 400MHz is for main rotational isomer): 7.7-7.64 (m, 2H), 7.44-7.3 (m, 3H), 7.15 (s, 1H), 4.88-4.78 (m, 1H), 4.06-4.0 (m, 1H), 3.46-3.4 (m, 1H), 2.06-1,4 (m), 1.24 (d, J=6.7,3H), 1.057 (d, J=6.9,3H), 1.01 (d, J=6.7.3H).
Step IV
As described in embodiment 25 steps 7, with the 3-[sec.-propyl--(trans-5-methyl-tetrahydrochysene-pyrans-the 2-carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (15mg, 0.038mmol) change 3-[sec.-propyl-(trans-5-methyl-tetrahydrochysene-pyrans-the 2-carbonyl)-amino into]-5-phenyl-thiophene-2-carboxylic acid (compound 27) (10mg, 68%).NMR
1H (CD
3OD, 400MHz is for main rotational isomer): 7.7-7.64 (m, 2H), 7.44-7.3 (m, 3H), 7.142 (s, 1H), 5.0-4.75 (m), 3.9-3.65 (m), 2.63 (t), 2.0-1,4 (m), 1.24 (d, 3H), 1.07 (d, 3H), 0.67 (d, 3H).
Embodiment 28
3-[(is trans-4-methyl-cyclohexyl alkyl carbonyl)-(tetrahydrochysene-thiapyran-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid compound 34,3-[(1,1-dioxy-tetrahydrochysene-thiapyran-4-yl)-(trans-4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid compound 37 and step VII:3-[(be trans-4-methyl-cyclohexyl alkyl carbonyl)-(1-oxygen-tetrahydrochysene-thiapyran-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid compound 68.
Step I
As described in embodiment 13 steps 1, adopt Bu
2SnCl
2(60.5mg, 0.2mmol) and PhSiH
3(0.476g, 0.542mL), in THF (1.0mL), make 3-amino-5-phenyl-thiophene-2-carboxylic acid methyl esters (0.933g, 4.0mmol) and tetrahydrochysene-thiapyran-4-ketone (0.464g, 4.0mmol) reduction amination, obtain 5-phenyl-3-(tetrahydrochysene-thiapyran-4-base is amino)-thiophene-2-carboxylic acid methyl esters (0.753g, 56.3%).NMR
1H(CDCl
3，400MHz)：7.64-7.6(m，2H)，7.44-7.34(m，3H)，6.9(brm，1H)，6.81(brs，1H)，3.84(s，3H)，3.5-3.4(m，1H)，2.85-2.7(m，4H)，2,4-2.25(m，2H)，1.85-1.7(m，2H)。
Step II
Described according to embodiment 19 steps 2, to 5-phenyl-3-(tetrahydrochysene-thiapyran-4-base is amino)-thiophene-2-carboxylic acid methyl esters (0.2g, 0.6mmol) and 4-methyl-cyclohexyl alkane dicarbonyl chloride carry out amidation, obtain 3-[(trans-4-methyl-cyclohexyl alkyl carbonyl)-(tetrahydrochysene-thiapyran-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (0.208g, 75.7%).NMR
1H(CDCl
3，400MHz)：7.68-7.62(m，2H)，7.5-7.4(m，3H)，7.04(s，1H)，4.68-4.58(m，1H)，3.86(s，3H)，2.9-1.2(m，16H)，0.78(d，3H)，0.76-0.56(m，2H)。
Step II I
Described according to embodiment 24 steps 3, with LiOH to 3-[(trans-4-methyl-cyclohexyl alkyl carbonyl)-(tetrahydrochysene-thiapyran-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (60mg, 0.13mmol) be hydrolyzed, obtain 3-[(trans-4-methyl-cyclohexyl alkyl carbonyl)-(tetrahydrochysene-thiapyran-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid (compound 34) (38mg, 65.9%).NMR
1H(CD
3OD，400MHz)：7.76-7.72(m，2H)，7.48-7.38(m，3H)，7.34(s，1H)，4.52-4.42(brt，1H)，2.9-2.5(m，4H)，1.8-1.2(m，9H)，0.773(d，J＝6.4.3H)，0.76-0.56(m，2H)。
Step IV
3-[(in step 2 is trans-4-methyl-cyclohexyl alkyl carbonyl)-(tetrahydrochysene-thiapyran-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (119mg, 0.26mmol) in the ice-cooled solution of DCM (1.0mL), a collection of adding m-chlorine peroxybenzoic acid (90mg, 60%, 0.312mmol), stirred 1 hour.Reaction mixture dilutes with DCM, uses NaHCO
3Saturated aqueous solution, the salt water washing, dry and concentrated.Resistates is on preparation type TLC, use the 50%EtOAc-hexane to carry out purifying, obtain 3-[(1,1-dioxy-tetrahydrochysene-thiapyran-4-yl as elutriant)-(trans-4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (88mg, 69%), is white solid.NMR
1H(CDCl
3，400MHz)：7.68-7.6(m，2H)，7.5-7.4(m，3H)，7.03(s，1H)，4.96-4.84(m，IH)，3.86(s，3H)，3.28-2.94(m，4H)，2.36-1.2(m，11H)，0.776(d，J＝4.8，3H)，0.76-0.54(m，2H)。
Step V
Described according to embodiment 25 steps 7, with LiOH hydrolysis 3-[(1,1-dioxy-tetrahydrochysene-thiapyran-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (47mg, 0.095mmol), obtain 3-[(1,1-dioxy-tetrahydrochysene-thiapyran-4-yl)-(trans-4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid (compound 37) (38mg, 84%).NMR
1H(CD
3OD，400MHz)：7.697(d，J＝7.17.2H)，7.426(t，2H)，7.35(t，1H)，7.23(s，1H)，4.72(brt，1H)，3.4-3.26(m，2H)，3.3-2.54(m，2H)，2.48-2.14(m，4H)，1.96-1.2(m，8H)，0.76-0.56(m，2H)，0.776(d，J＝6.6，3H)。
Step VI
3-[(in step 2 is trans-4-methyl-cyclohexyl alkyl carbonyl)-(tetrahydrochysene-thiapyran-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (57mg, 0.124mmol) EtOH (1.2mL) stirred solution in, a collection of adding monoperphthalic acid magnesium (29.6mg, 0.06mmol), stirred 24 hours.This reaction mixture dilute with water extracts with EtOAc.The organic solution salt water washing that merges, drying, and concentrate.Resistates uses 5%MeOH-DCM to carry out purifying on preparation type TLC, obtain 3-[(trans-4-methyl-cyclohexyl alkyl carbonyl)-(1-oxygen-tetrahydric thiapyran-4-group)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (30mg, 51%).NMR
1H (CDCl
3, 400MHz is to main isomer): 7.66-7.6 (m, 2H), 7.5-7.4 (m, 3H), 7.09 (s, 1H), 4.84-4.76 (t, 1H), 3.85 (s, 3H), 3.4-1.2 (m), 0.772 (d, J=6.6,3H), 0.74-0.56 (m, 2H).
Step VII
Described according to embodiment 25 steps 7, with LiOH hydrolysis 3-[(trans-4-methyl-cyclohexyl alkyl carbonyl)-(1-oxygen-tetrahydric thiapyran-4-group)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (30mg, 0.063mmol), obtain 3-[(trans-4-methyl-cyclohexyl alkyl carbonyl)-(1-oxygen-tetrahydric thiapyran-4-group)-amino]-5-phenyl-thiophene-2-carboxylic acid (compound 68) (15mg, 51.8%).NMR
1H (CD
3OD, 400MHz is to main isomer): 7.76-7.7 (m, 2H), 7.5-7.38 (m, 3H), 7.39 (s, 1H), 4.74-4.56 (m, 1H), 3.5-1.2 (m), 0.782 (d, J=6.4,1H), 0.75-0.55 (m, 2H).
Embodiment 29
5-(4-chloro-phenyl)-3-[(4-methyl-cyclohexyl alkyl carbonyl)-(1-methyl-piperidin-4-yl)-amino]-thiophene-2-carboxylic acid compound 57
Step I
Described according to embodiment 19 step I, adopt Bu
2SnCl
2(0.598g, 1.92mmol) and PhSiH
3(2.58mL, 21.6mmol), in THF (4.6mL) to 3-amino-thiophene-2-carboxylic acid methyl esters (3.0g, 19.1mmol) and 4-oxygen-piperidines-1-carboxylic acid benzyl ester (4.46g, 19.1mmol) carry out reduction amination, obtain 4-(2-methoxycarbonyl-benzene sulphur-3-base is amino)-piperidines-1-carboxylic acid benzyl ester (7.25g, quantitative).NMR
1H(CDCl
3，400MHz)：7.4-7.3(m，6H)，6.9-6.78(m，1H)，6.7-6.6(m，1H)，5.14(s，2H)，4.15-4.0(m，2H)，3.81(s，3H)，3.6-3.45(m)，3.1(brt，2H)，2.1-1.9(m，2H)，1.6-1.45(m，2H)。
Step II
According to embodiment 19, Step II is described, to 4-(2-methoxycarbonyl-benzene sulphur-3-base is amino)-piperidines-1-carboxylic acid benzyl ester (3.7g, 10mmol) carry out amidation with cyclohexyl chloride, obtain 4-[(2-methoxycarbonyl-benzene sulphur-3-yl)-(trans-4-methyl-cyclohexyl alkyl carbonyl)-amino]-piperidines-1-carboxylic acid benzyl ester (3.0g, 60%).NMR
1H(CDCl
3，400MHz)：7.55(d，1H)，7.36-7.26(m，5H)，6.82(d，1H)，5.05(brs，2H)，4.82-4.7(m，1H)，4.31-4.1(m，2H)，3.82(s，3H)，2.8-2.75(m，2H)，1.9-0.9(m，11H)，0.78(d，3H)，0.74-0.5(m，2H)。
Step II I
At 40psi, black with Pd/ to 4-[(2-methoxycarbonyl-benzene sulphur-3-yl)-(trans-4-methyl-cyclohexyl alkyl carbonyl)-amino]-piperidines-1-carboxylic acid benzyl ester (3.0g, 6.02mmol) carried out hydrogenation 36 hours, obtain 3-[(trans-4-methyl-cyclohexyl alkyl carbonyl)-piperidin-4-yl-amino]-thiophene-2-carboxylic acid methyl esters (1.0g, 45. 6%).NMR
1H(CDCl
3，400MHz)：7.525(d，J＝5.3，1H)，6.84(d，J＝5.3，1H)，4.674(tt，1H)，3.81(s，3H)，3.1-2.94(m，2H)，2.74-2.6(m，2H)，2.26-2.2(m，1H)，1.9-1.0(m，11H)，0.75(d，J＝6.6，3H)，0.7-0.5(m，2H)。
Step IV
3-[(in step 3 is trans-4-methyl-cyclohexyl alkyl carbonyl)-piperidin-4-yl-amino]-thiophene-2-carboxylic acid methyl esters (1.0g, 2.7mmol) 1, in the stirred solution of 2-ethylene dichloride (10mL), the order equal portions add the 37%HCHO aqueous solution (0.45mL, 5.4mmol) and NaBH (OAC)
3(2.86g, 13.5mmol), stirring is spent the night, and with this reaction of going out suddenly of the equivalent 10%NaOH aqueous solution, extracts with DCM.The organic extract liquid salt water washing that merges, drying, and concentrate acquisition 3-[(trans-4-methyl-cyclohexyl alkyl carbonyl)-(1-methyl-piperidin-4-yl)-amino]-thiophene-2-carboxylic acid methyl esters (0.876g, 85.5%), be white solid.NMR
1H(CDCl
3，400MHz)：7.54(d，1H)，6.86(d，1H)，4.7-4.6(m，1H)，3.85(s，3H)，2.9-2.7(m，2H)，2.22(s，3H)，2.15-1.1(m，14H)，0.8(d，3H)，0.75-0.5(m，2H)。
Step V
(0.3mL, (ethereal solution of 2.0M, 1.0mL 2.0mmol), stirred 24 hours to add positive BuMgCl in THF 2.14mmol) (10mL) stirred solution at diisopropylamine.In the solution that forms, drip 3-[(trans-4-methyl-cyclohexyl alkyl carbonyl)-(1-methyl-piperidin-4-yl)-amino]-thiophene-2-carboxylic acid methyl esters (0.189g, THF 0.5mmol) (2.0mL) solution, stirring at room 1 hour.(1.28g, THF 5.0mmol) (2.0mL) solution stirred 1 hour to add iodine then.Use 10%Na then
2S
2O
3The aqueous solution this reaction of going out suddenly with the EtOAc extraction, use the salt water washing, drying, and concentrated.Resistates is used DCM/CHCl combining on the silica gel paxillae of elutriant
3/ MeOH/Et
3N (200: 90: 16: 1) carry out purifying as elutriant, obtain 5-iodo-3-[(trans-4-methyl-cyclohexyl alkyl carbonyl)-(1-methyl-piperidin-4-yl)-amino]-thiophene-2-carboxylic acid methyl esters (0.250g, quantitatively).NMR
1H(CDCl
3，400MHz)：7.05(s，1H)，4.68-4.55(m，1H)，3.83(s，3H)，2.95-2.8(m，2H)，2.26(s，3H)，2.2-1.1(m，14H)，0.819(d，J＝6.3.3H)，0.75-0.6(m，2H)。
Step VI
At 4-chlorophenylboronic acid (46.9mg, 0.3mmol) and the 5-iodo-3-[(of step 5 trans-4-methyl-cyclohexyl alkyl carbonyl)-(1-methyl-piperidin-4-yl)-amino]-(50mg adds Pd (PPh in toluene/MeOH of 5: 1 (2.0mL) mixture 0.099mmol) to the thiophene-2-carboxylic acid methyl esters
3)
4(12.0mg, 0.01mmol 10mol%) at the solution of toluene (1.0mL), add 2M Na subsequently
2CO
3The aqueous solution (0.1mL, 0.2mmol).70 ℃ of reaction mixtures that add thermosetting 16 hours are cooled to room temperature, pass through MgSO
4Filter, and wash with EtOAc.Evaporating solvent, resistates is used DCM/CHCl on preparation type TLC (1mm, 60A °)
3/ MeOH/Et
3N (100: 90: 16:, provide 5-(4-chloro-phenyl)-3-((trans-4-methyl-cyclohexyl alkyl carbonyl)-(1-methyl-piperidin-4-yl)-amino)-thiophene-2-carboxylic acid methyl esters (35.0mg, 71.5% productive rate) 1) for elutriant carries out purifying.NMR
1H(CDCl
3，400MHz)：7.53(d，J＝8.3，2H)，7.4(d，J＝8.5，2H)，7.0(s，1H)，4.67-4.58(m，1H)，3.84(s，3H)，2.82-2.64(m，1H)，2.2(s，3H)，2.14-1.35(m)，0.763(d，J＝6.6，3H)，0.76-0.56(m，2H)。
Step VII
Described according to embodiment 25 step VII, with LiOH hydrolysis 5-(4-chloro-phenyl)-3-[(trans-4-methyl-cyclohexyl alkyl carbonyl)-(1-methyl-piperidin-4-yl)-amino]-thiophene-2-carboxylic acid methyl esters (19mg, 0.039mmol), obtain 5-(4-chloro-phenyl)-3-[(4-methyl-cyclohexyl alkyl carbonyl)-(1-methyl-piperidin-4-yl)-amino]-thiophene-2-carboxylic acid (compound 57) (9.0mg, 48.6%).NMR?
1H(CD
3OD?400MHz)：7.761(d，J＝8.8，2H)，7.487(d，J＝8.5，2H)，7.476(s，1H)，4.7(t，1H)，3.59-3.49(m，2H)，3.2-3.11(m，2H)，2.81(s，3H)，2.3-1.2(m，12H)，0.791(d，J＝6.59，3H)，0.88-0.5(m，2H)。
Adopt similar step, preparation compound 45, compound 54, compound 55, compound 56 and compound 58.
Embodiment 30
3-[[1-(4-methoxyl group-benzyl)-2-oxygen-piperidin-4-yl]-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid
Compound 42.
PMB=is to methoxy-benzyl
Step I
In nitrogen, (anhydrous methanol 8.01mmol) (5ml) solution is cooled to 0 ℃ for 690mg, 722 μ L, and (MeOH 7.28mmol) (1.0ml) solution is handled for 951 μ l, 1.0g to drip metacrylate in 2 minutes with right-methoxybenzylamine.15.5 after hour, under atmospheric pressure distill this clear solution, remove MeOH.High boiling residue (＞170
-) formation 3-(4-methoxyl group-benzylamino)-methyl propionate (1.88g, quantitatively)
1H (300MHz, CDCl
3) 1.78 (bs, 1H), 2.45 and 2.53 (t, J=3.0Hz, 2H), 2.77 and 2.87 (t, J=3.0Hz, 2H), 3.46 and 3.67 (s, 3H), 3.64 and 3.73 (s, 3H), 3.88 (m, 2H), 6.78 and 6.85 (m, 2H), 7.17 and 7.23 (m, 2H).
Step II
With pure dimethyl malonate (7.4ml, 8.5g, 64mmol, 8eq) solution is heated to 170 ° in a flask, drips the solution of the dimethyl malonate (0.92ml) of 3-(4-methoxyl group-benzylamino)-methyl propionate in 40 minutes.169-170 ℃ of reacting by heating thing 1.5 hours, TLC show that initial amine completely consumed is low polar compound.During cooling, crude product is added on the silica column, uses CH earlier
2Cl
2Wash-out is removed excessive dimethyl malonate, uses (hexane: CH then
2Cl
2: wash-out EtOAc=1: 1: 1).Collect N-(4-methoxyl group-benzyl)-N-(2-methoxycarbonyl-ethyl)-propanedioic acid methyl esters (1.502g, 58%) of colorless oil; (300MHz, CDCl
3) 2.52-2.61 (t, J=6.0Hz, 2H), 3.62-3.70 (s, 3H), 3.65-3.74 (s, 3H), 3.77-3.78 (s, 3H), 4.50-4.54 (s, 2H), 6.82-6.88 (m, 2H), 7.68-7.18 (m, 2H).
Step II I
Moisture K
2CO
3(3.2g, 23.2mmol, 5eq) and 18-crown-6 (with benzene azeotropic for several times) (122mg, 0.464mmol, 10mol%) in dry toluene (4ml), mix, in nitrogen, be heated to backflow, in 40 minutes, drip N-(4-methoxyl group-benzyl)-N-(2-methoxycarbonyl-ethyl)-propanedioic acid methyl ester solution then and handle.Reflux after 7 hours, reactant water (4ml) and toluene (4ml) dilution are cooled to 0 ℃ then, and carefully are acidified to pH 1.7 with 0.1N HCl.Then, mixture CH
2Cl
2(3 * 80ml) extracted several times, the organism of merging is dry and be evaporated to brown oil (1.33g).This brown oil is handled with 10% oxalic acid aqueous solution, and reflux 6.5 hours.Mixture CH
2Cl
2Extraction repeatedly, the organism that merges is dry and flash to yellow oil (1.03g).Crude product is used (CH on silicagel column
2Cl
2: MeOH 30: 1) carry out purifying as elutriant, obtain 1-(4-methoxyl group-benzyl)-piperidines-2, the 4-diketone is light brown solid (750mg, 69%); (300MHz, CDCl
3) 2.52 (t, J=5.7Hz, 2H), 3.41 (s, 2H), 3.47 (t, J=5.7Hz, 2H), 3.80 (s, 3H), 4.62 (s, 2H), 6.16-6.88 (m, 2H), 7.20 (m, 2H).
Step IV
With 3-amino, (459mg, 1.96mmol) and 1-(4-methoxyl group-benzyl)-piperidines-2, (457mg, suspension 1.96mmol) be at 21 ℃, N for the 4-diketone for 5-phenyl thiophene 2-carboxylate methyl ester
2In, (29mg, 0.098mmol 5mol%) handle, and (2.15mmol 1.1eq) handles for 266 μ l, 233mg with phenyl silane after 5 minutes with the dichloride dibutyl tin.Stir this heterogeneous mixture 18 hours at 21 ℃, form clear solution.Continue this reaction 5 hours, be evaporated to thick oily matter (1.27g) then.Crude product is used (hexane: CH on silicon-dioxide
2Cl
2: EtOAc=1: 1: 1) as the elutriant purifying, provide 3-[1-(4-methoxyl group-benzyl)-2-oxygen-piperidin-4-yl amino]-(the 1-methyl-oneself is-1,3 years old for 5-, the 5-trialkenyl)-the thiophene-2-carboxylic acid methyl esters, be yellow foams (432mg, 49%) (300MHz, CDCl
3) 1.8-1.9 (m, 1H), 2.25-2.44 (m, 1H), 2.95 (dd, J=1.5Hz, J=3.90Hz, 1H), 3.98 (dd, J=1.5Hz, J=3.90Hz, 1H), 3.22-3.40 (m, 4H), 3.80 (s, 3H), 3.33 (s, 3H), 3.85-3.93 (m, 1H), 4.08 (m, 2H), 6.81 (s, 1H), 6.85-6.9 (m, 2H), 7.20-7.24 (m, 2H), and 7.36-7.42 (m, 3H), 7.59-7.61 (m, 2H).
Step V
With trans 4-methylcyclohexanecarboxylic acid (56mg, 0.399mmol, 1.2eq) 1,2-ethylene dichloride (1ml) solution, at 0 ℃, N
2Down, with the oxalyl chloride (CH of 2.0M
2Cl
2Solution) (231 μ l, 0.46mmol 1.4eq) handles, and (0.1mmol 30mol%) handles for 8 μ l, 7mg to use dimethyl formamide subsequently.After 1 hour, reaction is with 3-[1-(4-methoxyl group-benzyl)-2-oxygen-piperidin-4-yl amino]-5-(the 1-methyl-oneself-1,3, the 5-trialkenyl)-thiophene-2-carboxylic acid methyl esters (150mg, DCE 0.33mmol) (2ml) solution-treated.Then reactant is placed 90 ℃ of baths, making refluxes spent the night in 21 hours.Remove the solvent in the reactant, resistates (212mg) is used in conjunction with elution chromatography, with (hexane: CH
2Cl
2: EtOAc=1: 1: 1) carry out purifying as elutriant, 3-[[1-(4-methoxyl group-benzyl)-2-oxygen-piperidin-4-yl is provided]-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-(the 1-methyl-oneself-1,3, the 5-trialkenyl)-thiophene-2-carboxylic acid methyl esters (21mg, 11%), is yellow foams; (300MHz, CDCl
3) 0.55-0.73 (m, 1H), 0.77 (d, J=5.4Hz, 3H), 1.26-1.30 (m, 12H), 1.94-2.12 (m, 1H), 2.14-2.19 (m, 1H), 2.40 (dd, J=9.0Hz, J=12.0Hz, 1H), 2.64-2.70 and 2.80-2.84 (m, 1H), 3.10-3.15 (m, 3H), 3.79-3.82 (m, 1H), 3.78 (s, 3H), 3.83-3.87 (s, 3H), 4.32 (t, J=12.0Hz, 1H), 4.62 (dd, J=5.7Hz, J=10.8Hz, 1H) ' 4.50-5.0 (m, 1H), 6.79-6.83 (m, 2H), 6.99-7.14 (m, 2H), 7.26-7.48 (m, 3H), 7.61-7.65 (m, 2H).
Step VI
Described according to embodiment 25 step VII, with LiOH hydrolysis 3-[[1-(4-methoxyl group-benzyl)-2-oxygen-piperidin-4-yl]-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (40mg, 0.069mmol), 3-[[1-(4-methoxyl group-benzyl)-2-oxygen-piperidin-4-yl is provided]-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid (compound 42), be white solid (8.4mg, 21%); (300MHz, acetone-d
6) 0.42-0.62 (m, 1H), 0.64 (d, J=4.18Hz, 3H), 1.16-1.34 (m, 7H), 1,40-1.54 (m, 4H), 1.58-1.66 (m, 2H), 1.76-1.78 (m, 1H), 1.87-1.90 (m, 1H), 1.98-1.99 (m, 1H), 2.08-2.09 (m, 1H), 3.63 (s, 3H), 4.23 (dd, J=5.7Hz, J=12.0Hz, 1H), 4.44 (dd, J=1.5Hz, J=11.7Hz, 1H), 4.66-4.80 (m, 1H), 6.20-6.23 (m, 2H), 7.04-7.07 (m, 2H), 7.31-7.40 (m, 3H), 7.44-7.53 (s, 1H), 7.69-7.75 (m, 2H).
Embodiment 31
3-[(1-methylsulfonyl-piperidin-4-yl)-(4-methyl cyclohexane alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid
Compound 70.
The CBZ=benzyloxycarbonyl
Carry out step I-III according to the mode that is similar to embodiment 30.
Step IV
With the 3-[(1-piperidin-4-yl)-(4-methyl cyclohexane alkyl carbonyl)-amino]-(44mg, DCM 0.1mmol) (1.0ml, ca 0.1M) solution is at 21 ℃ and N for 5-phenyl-thiophene-2-carboxylic acid methyl esters
2Under use triethylamine (0.21mmol 2.1eq) handle for 29 μ l, 21mg, and (0.2mmol 1.2eq) handles for 15.5 μ l, 23mg to use methylsulfonyl chloride subsequently.Small amount of precipitate appears.1.5 after hour, finish reaction.With DCM diluted mixture thing, use 1N HCl successively, water, salt water washing, drying.The evaporation organic extract liquid produces colloid (56mg), on the silicon-dioxide in conjunction with elutriant, with (hexane: CH
2Cl
2: EtOAc=1: 1: 1) carry out purifying, obtain 3-[(1-methylsulfonyl-piperidin-4-yl as elutriant)-(4-methyl cyclohexane alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (40mg, 78%);
1H (300MHz, CDCl
3) 0.50-0.67 (m, 1H), 0.70 (d, J=4.8Hz, 3H), 1.16-1,43 (m, 6H), 1,48-1.64 (m, 10H), 1.80-1.86 (m, 1H), and 1.90-2.0 (m, 1H), 2.62-2.759 (m, 2H), 2.68 (s, 3H), and 3.68-3.75 (m, 1H), 3.76-3.82 (m, 1H), 3.85 (s, 3H), 4.61-4.71 (m, 1H), 6.94 (s, 1H), and 7.35-7.43 (m, 3H), 7.55-7.59 (m, 2H).
Step V
As previously described, with lithium hydroxide (2M, 114 μ L, 5.5mg, 0.23mmol) hydrolysis 3-[(1-methylsulfonyl-piperidin-4-yl)-(4-methyl cyclohexane alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (40mg 0.077mmol), obtains 3-[(1-methylsulfonyl-piperidin-4-yl after the acidifying)-(4-methylcyclohexane-carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid (29mg, 74%), is colourless powder;
1H (300MHz, MeOD) 0.56-0.72 (m, 1H), 0.79 (d, J=4.8Hz, 3H), 1.23-1,44 (m, 5H), 1.51-1.79 (m, 6H), 1.92-1.98 (m, 1H), 2.04-2.14 (m, 2H), 2.78-2.90 (m, 2H), 2.80 (s, 3H), 3.66-3.74 (m, 1H), 3.75-3.81 (m, 1H), 4.52-4.62 (m, 1H), 7.39 (s, 1H), and 7.40-7.90 (m, 3H), 7.73-7.77 (m, 2H).
Embodiment 32
3-[(2-amino-1-methyl-ethyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-(the 1-methyl-oneself-1,3, the 5-trialkenyl)-thiophene-2-carboxylic acid compound 36; 3-[(2-azido--1-methyl-ethyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-(the 1-methyl-oneself-1,3, the 5-trialkenyl)-thiophene-2-carboxylic acid compound 32
Step I
Will(S)-(+) 2-amino-1-propyl alcohol (1.04g, 13.85mmol), tert-butyldimethylsilyl chloride (2.09g, 13.85mmol) and triethylamine (2ml, mixture 1.05eq) stir in DCM and spend the night 16 hours.Dilute this reactant with DCM, water, saturated NH
4Cl, the salt water washing, dry and be evaporated to oily matter.Crude product uses 3%MeOH/CH at silica gel
2Cl
2Carry out purifying as elutriant, obtain 2-(tert-butyl-dimethyl-siloxy-)-1-methyl-ethamine (1.76g, 80%).
1H?NMR(300MHz，CDCl
3)0.0(s，6H)，0.82(s，9H)，0.99(d，J.＝6.5Hz，3H)，2.22(bs，2H)，2.98(bs，1H)，3.30(dd，J＝10Hz，J＝17.0Hz，1H)，3.48(dd，J＝10.0Hz，J＝4.3Hz，1H)。
Step II
(0.5g, toluene solution 1.6mmol) (10ml) be at 21 ℃, N for 3-bromine 5-phenyl thiophene 2-carboxylate methyl ester
2In with 2-(tert-butyl-dimethyl-siloxy-)-1-methyl-ethamine (301mg, 2.01mmol 1.2eq) handle, use subsequently acid chloride (38mg, 0.1eq), BINAP (105mg, 0.1eq) and CsCO3 (766mg, 1.4eq) processing.Reflux mixture 8 hours filters by Celite pad then.Wash Celite pad with EtOAc, washings dry and that evaporation merges becomes colloid, this colloid carries out purifying with the 3%EtOAc/ hexane as elutriant by the silicon-dioxide chromatography, obtain required compound 3-[2-(tert-butyl-dimethyl-siloxy-)-1-methyl-ethylamino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (645mg, 95%);
1H NMR (300MHz, CDCl
3) 0.0 (s, 6H), 0.82 (s, 9H), 1.24 (d, J=6.0Hz, 3H), 3.56-3.57 (m, 3H), 3.90 (s, 3H), 6.88 (bs, 1H), 6.84 (s, 1H), 7.30-7.40 (m, 3H), 7.56 (d, J=6.0Hz, 2H).
Step II I
At 21 ℃, N
2In to 3-[2-(tert-butyl-dimethyl-siloxy-)-1-methyl-ethylamino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (1.08g, 2.69mmol) MeOH (10ml) solution in, add pre-mixing at MeOH (100 μ L/1mL, 210 μ L 2.96mmol, 1.1eq) chloride solutions in.Carry out TLC after the reaction.Finish reaction, remove the solvent in the reactant, resistates silicon-dioxide with concentration raise gradually 5%, 20%, the 30%EtOAc/ hexane carries out purifying as elutriant then, and (the 1-methyl-oneself is-1 years old to obtain 3-(2-hydroxyl-1-methyl-ethylamino)-5-, 3, the 5-trialkenyl)-and the thiophene-2-carboxylic acid methyl esters, be yellow solid (518mg, 79%);
1H NMR (300MHz, CDCl
3) 1.25 (d, 6.6Hz, 3H), 3.52-3.62 (m, 1H), 3.73-3.77 (m, 2H), 3.84 (s, 3H), 6.92 (s, 1H), 7.36-7.42 (m, 3H), 3.62 (d, J=8.3Hz, 2H).
Step IV
Stir 3-(2-hydroxyl-1-methyl-ethylamino)-5-phenyl-thiophene-2-carboxylic acid methyl esters (213mg at 21 ℃; 0.257mmol); diazonium diethyl dicarboxylate (250uL, 1.59mmol, 2eq); diphenylphosphine acyl group nitride (343 μ L; 1.59mmol, 2eq) and triphenylphosphine (417mg, 1.59mmol; mixture 2eq) all consumes up to initial alcohol.The evaporation reaction thing is used the 5%EtOAc/ hexane to doing with rough resistates on biotage, carry out purifying with 100% toluene as elutriant subsequently.Separate obtaining 3-(2-azido--1-methyl-ethylamino)-5-phenyl-thiophene-2-carboxylic acid methyl esters, be solid (181mg, 78%);
1H NMR (300MHz, CDCl
3) 1.33 (d, J=6.6Hz, 3H), 3.38-3.46 (m, 2H), 3.78-3.81 (m, 1H), 6.82 (s, 1H), 7.36-7.41 (m, 3H), 7.60-7.62 (m, 2H).
Step V
Described according to embodiment 30 step V, trans 4-methylcyclohexanecarboxylic acid muriate (25mg with prepared fresh, 0.176mmol, 1.2eq) processing 3-(2-azido--1-methyl-ethylamino)-5-phenyl-thiophene-2-carboxylic acid methyl esters (60mg, 0.188mmol) solution, obtain 3-[(2-azido--1-methyl-ethyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (14.6mg, 17%);
1H NMR (300MHz, CDCl
3) 0.5-0.7 (m, 4H), 0.71 (d, J=6.6Hz, 3H), 1.23 (d, J=7.0Hz, 3H), 1,2-1,42 (m, 4H), 2.12-2.25 (m, 1H), 3.24 (dd,, J=5.6Hz, J=5.7Hz, 1H), 3.52 (dd, J=5.6Hz, J=5.7Hz, 1H), 3.80 (s, 3H), 4.82-4.90 (m, 1H), 7.20 (s, 1H), and 7.32-7.42 (m, 3H), 7.56-7.61 (m, 2H).
Step VI
With 3-[(2-azido--1-methyl-ethyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (14mg, 0.032mmol) De diox: water=4: 1.0.5ml) solution is according to embodiment 25, step 7 is described, with LiOH (4mg, 3eq) handle, obtain 3-[(2-azido--1-methyl-ethyl after the acidifying)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid (compound 32), be light green foams (11.2mg, 82%);
1H NMR (300MHz, acetone-d
6) 0.43-0.62 and 0.77-0.86 (m, 1H), 0.63-0.75 (d, J=5.1Ez, 3H), and 0.93-1.20 (d, J=5.2Hz, 3H), 1.78-1.85 and 1.97-2.10 (m, 2H), 3.20-3.68 (m, 1H), 3.32-3.50 (m, 1H), 4.40-4.50 (m, 1H), 7.3-7.4 (m, 3H), 7.44 (s, 1H), and 7.67-7.69 (m, 2H).
Step VII
3-[(2-azido--1-methyl-ethyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-((4mg 50%w/w) handles, at H 5-phenyl-thiophene-2-carboxylic acid with 10%Pd/C at 21 ℃ for 8mg, EtOH 0.19mmol) (0.2ml) solution
2Stirred 1.5 hours under the atmosphere.Pass through Celite pad with hot EtOAc filter reaction mixture, merging filtrate and washings, dry and evaporation becomes vitreum, obtains 3-[(2-amino-1-methyl-ethyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid (compound 36).(7mg, %);
1HNMR (300MHz, acetone-d
6) 0.43-0.65 and 0.75-0.90 (m, 4H), 1.17-1.64 (m, 6H), 1.81-1.90 and 2.01-2.40 (m, 3H), 2.54 (bs, 1H), 3.00 (bs, 1H), 3.50 and 3.70 (bs, 1H), 7.20 (s, 1H), 7.22-7.25 and 7.29-7.34 (m, 3H), 7.58-7.62 (m, 2H).
Same preparation compound 43, compound 20, compound 19, compound 18, compound 7 and compound 8.
Embodiment 33
3-[(1-cyano group-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid compound 77.
Step I
With 3-[(4-methyl-cyclohexyl alkyl carbonyl)-piperidin-4-yl-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (197mg, CH 0.45mmol)
2Cl
2(4.5mL) solution K
2CO
3(93mg, 0.67mmol) and cyanogen bromide (100mg 0.94mmol) handles.Stirring at room reaction mixture 1 hour, reflux 18 hours.Mixture at room temperature cools off, diatomite filtration.Filtrate is used AcOH (1N) and salt water washing, dry (Na
2SO
4) and concentrate.Resistates is by purification by silica gel column chromatography, with (2%MeOH/CH
2Cl
2), obtain 3-[(1-cyano group-piperidin-4-yl)-(4-methyl cyclohexane alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (154mg, 74% productive rate), be the weak yellow foam body.
Step II
With 3-[(1-cyano group-piperidin-4-yl)-(4-methyl hexanaphthene carbonyl)-amino]-(150mg 0.32mmol) is dissolved in 4: 1 De diox: H to 5-phenyl-thiophene-2-carboxylic acid methyl esters
2In the O mixture (3.2ml), and use LiOHH
2(20mg 0.48mmol) handles O.50 ℃ were stirred after 2 hours, removed and desolvated, and were distributed in the H that 5ml is acidified to pH 4 then
2Between O and the 5ml EtOAc.Separate organic layer and water ethyl acetate (2 * 5mL) washed twice.With the ethyl acetate layer drying (Na that merges
2SO
4) and concentrate.Resistates uses (5%MeOH/CH by purification by silica gel column chromatography
2Cl
2), obtain 3-[(1-cyano group-piperidin-4-yl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid (114.3mg, 78% productive rate), be the light green foams.
1HNMR(DMSO-d
6，400MHz)：7.80(m，2H)，7.45(m，4H)，4.44(m，1H)，3.35(m，2H)，3.13(m，2H)，1.96(t，1H)，1.88(d，1H)，1.75(m，1H)，1.70-1.40(m，6H)，1.20(m，3H)，0.70(d，3H)，0.60(m，2H)。
Embodiment 34
Cis-3-[(4-hydroxy-4-methyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid compound 86.
Step I
At-40 ℃, under the nitrogen atmosphere, to zinc powder (2.87g, 44.0mmol) and ethylene dibromide (1.00mL, 14.4mmol) in the stirred suspension of tetrahydrofuran (THF) (20mL), add titanium tetrachloride (10mL, solution in the methylene dichloride of 1M, 10mmol).Mixture heating up is to room temperature, and stirs 2 days under this temperature.At 3-[(4-methyl-cyclohexyl alkyl carbonyl)-(4-oxygen-cyclohexyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (0.200g, 0.440mmol, add this methylating reagent (2.5eq) in methylene dichloride 1eq) (2mL) solution, the mixture of generation was stirring at room 3 hours.Add saturated solution of sodium bicarbonate then, (3 * 30mL) extract reaction mixture with methylene dichloride.Merge organic phase, use dried over sodium sulfate, and concentrate.Crude product obtains the 3-[(4-methyl-cyclohexyl alkyl carbonyl of 160mg (81%) by chromatography (30% ethyl acetate/hexane) purifying)-(4-methylene radical-cyclohexyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters, be white solid.NMR
1H(CDCl
3，400MHz)：7.63?ppm(d，2H)；7.40ppm(m，3H)；6,98?ppm(s，1H)；4.82?ppm(tt，1H)；4.78?ppm(d，2H)；3.85?ppm(s，3H)；2.20?ppm(m，4H)；2.05?ppm(m，2H)；1.90?ppm(d，1H)；1.65?ppm(m，4H)；1,42ppm(m，1H)；1.30ppm(m，2H)；1.00ppm(m，2H)；0.78ppm(d，3H)；0.64ppm(m，2H)。
Step II
Under the room temperature, in water (1mL) and tetrahydrofuran (THF) (1mL) solution, add mercuric acetate (83.0mg, 0.277mmol, 1eq).Stir after 10 minutes, yellow solution be cooled to 0 ℃, drip solution 3-[(4-methyl-cyclohexyl alkyl carbonyl)-(4-methylene radical-cyclohexyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (125mg, 0.277mmol, 1eq).The mixture of 0 ℃ of stirring formation 1 hour.Add NaOH 3M (1mL) then, add again sodium borohydride (10.0mg, 0.277mmol, 1eq), this reaction mixture of stirring at room 15 minutes.(3 * 30mL) extract reaction mixture with methylene dichloride.Merge organic phase, use dried over sodium sulfate, and concentrate.Crude product is by chromatography (50% ethyl acetate/hexane) purifying, separate diastereomer, obtain 90mg cis-3-[(4-hydroxy-4-methyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters and 6.5mg be trans-3-[(4-hydroxy-4-methyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (74%), all be white solid.Cis-3-[(4-hydroxy-4-methyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters.NMR
1H(CDCl
3，400MHz)：7.63ppm(d，2H)；7.40?ppm(m，3H)；6,98?ppm(s，1H)；4.50?ppm(tt，1H)；3.85?ppm(s，3H)；2.00?ppm(m，1H)；1.80-1.20?ppm(m，15H)；1.18?ppm(s，3H)；0.78?ppm(d，3H)；0.64ppm(m，2H)。
Step II I
At cis-3-[(4-hydroxy-4-methyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid methyl esters (70.0mg, 0.149mmol, tetrahydrofuran (THF) 1eq) (1mL), add lithium hydroxide (19.0mg in water (0.5mL) and methyl alcohol (0.5mL) solution, 0.447mmol, 3eq).The mixture that stirring at room forms 3 hours is used ether (2 * 10mL) extractions then.Water phase separated discards the organic phase of merging.The acidifying water is to pH1, with methylene dichloride (3 * 30mL) extractions.The combined dichloromethane organic phase is with dried over sodium sulfate and concentrated.Crude product obtains cis-3-[(4-hydroxy-4-methyl-cyclohexyl of 50mg (74%) by chromatography purification (10% ethanol/methylene))-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid.NMR
1H(CDCl
3，400MHz)：7.60?ppm(d，2H)；7.39?ppm(m，3H)；7.03?ppm(s，1H)；4.51?ppm(bs，2H)；2.00?ppm(m，1H)；1.80-1.20?ppm(m，15H)；1.12?ppm(s，3H)；0.71ppm(d，3H)；0.60?ppm(m，2H)。
Embodiment 35
The sodium salt of the following compound of preparation
With 1: 1 diox/aqueous solution of carboxylic acid cpd A (1mmol), (1mmol 1eq) handles with 0.1 N NaOH solution at 0 ℃.Reaction stirred 15 minutes.Concentrate this solution then, and freeze-drying, sodium salt obtained; Carboxylic acid cpd B is a solid.
Embodiment 36
Compound is tabulated with relevant polymerase activity
Table 1
+++IC
50＜5μm
++IC
50?5-20μm
+IC
50＞20μm
Embodiment 37
In the RNA polymerase test that HCV RNA-relies on, estimate all compounds.
It is for referencial use that following reference is all included this paper in:
1.Behrens，S.，Tomei，L.，De?Francesco，R.(1996)EMBO?15.12-22
2.Harlow，E，and?Lane，D.(1988)Antibodies：A?Laboratory?Manual.ColdSpring?Harbord?Laboratory.Cold?Spring?Harbord.NY.
3.Lohmann，V.，Korner，F.，Herian，U.，and?Bartenschlager，R.(1997)J.Virol.71；，8416-8428
4.Tomei，L.，Failla，C.，Santolini，E.，De?Francesco，R.，and?La?Monica，N.(1993)J?Virol?67；4017-4026
5.US it is for referencial use that temporary patent application 10/166,031 is also included this paper in.
Employing contains RNA polymerase (NS5B albumen) the polymerization in vitro enzyme test of the recombinant HCV RNA-dependence of purifying, assessing compound.With recombinant baculovirus as carrier, at expressed in insect cells HCV NS5B.Describe below and be used for the clone, expression and the proteic experimental technique of purifying HCV NS5B.Describe the RNA polymerase test of the RNA-dependence of all compounds of test below in detail.
The expression of HCV NS5B albumen in insect cell: the proteic cDNA of NS5B of the whole HCV-Bk chain of will encoding, genotype 1b, by pcr amplification, the primer is
NS5Nhe 5 ' (5 '-
GCTAGCGCTAGCTCAATGTCCTACACATGG-3 ') and
XhoNS53 ' (5 '-
CTCGAGCTCGAGCGTCCATCGGTTGGGGAG-3 '), use plasmid pCD 3.8-9.4 as template (Tomei et al, 1993).NS5Nhe5 ' and XhoNS53 ' contain two NheI and XhoI site (underscore sequence) at its 5 ' end respectively.The dna fragmentation of amplification is cloned among the bacterial expression plasmid pET-21b (Novagen), between restriction site NheI and XhoI, produces plasmid pET/NS5B.This plasmid comes the coding region of PCR-amplification NS5B then as template, the primer is
NS5B-H9 (5 '-ATACATATGGCTAGCATGTCAATGTCCTACACATGG-3 ') and
NS5B-R4(5’-
GGATCCGGATCCCGTTCATCGGTTGGGGAG-3’)。NS5B-H9 crosses over 15 Nucleotide zones of plasmid pET-21b, will translate 8 Nucleotide (nt.7590-7607 is in the HCV of accession number M58335 sequence) that start codon (ATG) and hold corresponding to NS5B coding region 5 ' subsequently.NS5B-R4 contains two BamHI sites (underscore), is 18 Nucleotide (nt.9365-9347) corresponding to HCV genome terminator codon peripheral region subsequently.The extension increasing sequence of 1.8kb is digested with NheI and BamHI, and be connected in the pBlueBacII plasmid (Invitrogen) of digestion in advance.The recombinant plasmid called after pBac/NS5B that produces.The Sf9 cell carries out as the described scheme of manufacturers with the linearized baculovirus dna (Invitrogen) of the pBac/NS5B of 3 μ g and 1 μ g cotransfection together.Behind two-wheeled plaque purifying, separate obtaining NS5B-recombinant baculovirus, BacNSSB.(Harlow and Lane, 1988) are analyzed in the immunity printing and dyeing of the Sf9 cell that infects by BacNS5B-, adopt the proteic rabbit polyclonal antiserum of NS5B that contains resistance propylhomoserin tail at escherichia coli expression (anti--as NS5B), to measure and have reorganization NS5B albumen.In 1 liter of rotary flask with this through the virus infection Sf9 of plaque purifying cell, cell density is 1.2 * 10
6Cell/ml, infection multiplicity is 5.
Press top described preparation solubility reorganization NS5B albumen with the Sf9 cell that infects.Infect after 60 hours, collecting cell is also used phosphate-buffered saline (PBS) washed twice.As described in (1997) such as Lohmann, done some improvement, the dissolving all proteins.Briefly, at three step S1, S2, among the S3, with lysis buffer (LB) I, LBII and LBIII extract protein (Lohmann et al, 1997).The composition of revising LBII makes and contains 0.1% triton x-100 and 150mM NaCl, to reduce the proteinic amount of dissolving NS5B in this step.In addition, in the whole proposal process, avoid using the ultrasonication cell extract, to preserve the integrity of protein structure.
Adopt fast protein liquid chromatography (FPLC) purification of Recombinant NS5B:
Solubility NS5B protein in the dilution S3 component reduces NaCl concentration to 300mM, cultivates 2 hours for 4 ℃ with DEAE sepharose 4B (Amersham-Pharmacia) then, as described in Behrens et al. (1996) in batches.Adopt SW41 rotary head (Beckman), 4 ℃, centrifugal 15 minutes of 25000rpm removes unconjugated material.Further the dilution supernatant liquor is reduced to 200mM with NaCl concentration, is added to 5ml HiTrap with the 1ml/min flow velocity subsequently
On the heparin column (Amersham-Pharmacia), this post is connected in FPLC
System (Amersham-Pharmacia).Adopt the N continuous aCl gradient 25ml volume of 0.2-1M, bonded protein is eluted in the 1ml component.The component that contains NS5B, is identified with the anti--NS5B antiserum(antisera) of dilution in 1: 2000 then through immunoblotting with SDS-PAGE (SDS-PAGE).Merge positive component, with PD-10 post (Amersham-Pharmacia), with 50mM NaPO
4PH 7.0,20% glycerine, 0.5% triton x-100 and the agent of 10mM DTT exchange elution buffer.With the 0.1ml/min flow velocity sample is added to 1ml HiTrap then
On the SP post (Amersham-Pharmacia).Bonded protein adopts 0-1M N continuous aCl gradient, and the 15ml volume carries out wash-out.The component of wash-out SDS-PAGE and immunoblotting analysis.Perhaps, described according to manufacturers, with Silver Stain Plus test kit (BioRad), observe these protein by the dyeing of the silver behind the SDS-PAGE.Measure the RdRp activity (referring to following) of positive component, merge active maximum component ,-70 ℃ of storages in 40% glycerine solution.
Be used to estimate external HCV RdRp liquid sudden strain of a muscle plate (Flashplate) the approaching test of flicker (STREP-FLASH ASSAY) of congener:
This test comprises: measure and be trapped in bag by the flicker microtitration liquid sudden strain of a muscle plate of Streptomycin sulphate avidin
TMMix in (NENLife Science Products inc, MA, USA, SMP 103A) lip-deep polyrA/ biotinylated oligonucleotide dT template-primer [
3H] amount of labelled with radioisotope UTP.Briefly, with the 5 ' vitamin H-oligonucleotide dT of 400ng/ μ l polyrA solution (Amersham Pharmacia Biotech) with 20pmol/ μ l
15By volume mix.95 ℃ of sex change templates and primer 5 minutes were cultivated 10 minutes for 37 ℃ then.With annealed template-primer with the dilution of Tris-HCl damping fluid, make its in conjunction with bag by the liquid of Streptomycin sulphate avidin dodge plate and spend the night.Discard unconjugated material, add the 10 μ l solution that contain compound, add 10 μ l subsequently and contain 50mM MgCl
2, 100mM Tris-HCl pH 7.5, the solution of 250mM NaCl and 5mM DTT.Adding solution that 30 μ l contain enzyme and substrate reaches following concentration and causes enzymatic reaction: 25 μ MUTP, 1 μ Ci[
3H] UTP and 100nM recombinant HCV NS5B.Room temperature is carried out RdRp reaction 2 hours, and the 0.15M NaCl solution with 250 μ l washes each Kong Sanci then, and 37 ℃ of dry airs are with liquid scintillation counter (Wallac Microbeta Trilex, Perkin-Elmer, MA, USA) counting.The results are shown in Table 1.
Adopt the above embodiment of the present invention general introduction and specifically described reactant and/or operational condition to replace, can successfully repeat the example of front equally.
By noted earlier, those skilled in the art are not difficult to determine essential characteristic of the present invention, and are not breaking away under design of the present invention and the scope, can make various changes and modification to the present invention, make the present invention be adapted to various uses and condition.
Claims (46)
1. the compound of structural formula below a kind, or its pharmacy acceptable salt;
In the formula:
Z is a cyclohexyl, is not substituted or is selected from following substituting group and replaces by one or more: oxygen base, halogen, SO
2Rf, CONRgRh, C
1-6Alkyl, C
6-12Aralkyl, C
6-12Aryl, C
1-6Alkoxyl group, C (O) C
1-6Alkyl, C
3-10Heterocycle, hydroxyl, NRgRh, C (O) ORf or cyano group;
Y is a phenyl, is not substituted or is selected from following substituting group and replaces by one or more: halogen, nitro, SO
2Rf, C
1-6Alkyl, C
1-6Alkoxyl group, C (O) C
1-6Alkyl, C (O) ORf, cyano group, azido-;
Rf wherein, Rg and Rh are H or C independently of one another
1-6Alkyl;
X is a cyclohexyl, is not substituted or is selected from following substituting group and replaces by one or more: C
1-6Alkyl, hydroxyl, halogen, C
2-6Thiazolinyl, C
2-6Alkynyl or C
1-6Alkoxyl group;
M is 0;
When Y was unsubstituted phenyl, X was not the 4-methylcyclohexane.
2. compound as claimed in claim 1 is characterized in that, X is 4-methyl-cyclohexyl base or 2-hydroxy-4-methyl-cyclohexyl.
3. compound as claimed in claim 1 is characterized in that Y is a phenyl.
4. as each described compound among the claim 1-3, it is characterized in that described pharmacy acceptable salt is a sodium salt.
5. each described compound is used for the treatment of or prevents application in the medicine that host's flavivirus infects in preparation among the claim 1-4.
6. application as claimed in claim 5; described medicine also comprises at least a other the following preparations that are selected from: virus serine protease class inhibitor; viral polymerase inhibitors; virus helicase inhibitor; immunomodulator, antioxidant, antiseptic-germicide; therapeutic vaccine, hepatoprotective or antisense agents.
7. application as claimed in claim 5 is characterized in that, described medicine also comprises at least a other the following preparations that are selected from: interferon alpha, virazole; Silymarin, il-1 2, amantadine; rnase, thymosin, N-acetylcysteine or ciclosporin.
8. application as claimed in claim 5 is characterized in that it is infection with hepatitis C virus that described flavivirus infects.
9. be used for suppressing or reduce application in the medicine of host's flavivirus polymerase activity in preparation as each described compound among the claim 1-4.
10. application as claimed in claim 9 is characterized in that, described polysaccharase is the RNA-polysaccharase that RNA-relies on.
11. application as claimed in claim 9 is characterized in that, described polysaccharase is the hepatitis C virus polysaccharase.
12. a pharmaceutical composition is characterized in that, said composition comprises at least a as each described compound among the claim 1-4 and at least a pharmaceutically acceptable vehicle or vehicle.
13. pharmaceutical composition as claimed in claim 12; described pharmaceutical composition also comprises at least a other the following preparations that are selected from: virus serine protease class inhibitor; viral polymerase inhibitors; virus helicase inhibitor; immunomodulator, antioxidant, antiseptic-germicide; therapeutic vaccine, hepatoprotective or antisense agents.
14. pharmaceutical composition as claimed in claim 12; it is characterized in that; described pharmaceutical composition also comprises at least a other the following preparations that are selected from: interferon alpha; virazole, Silymarin, il-1 2; amantadine; rnase, thymosin, N-acetylcysteine or ciclosporin.
15. compound as claimed in claim 1, wherein Z is oxygen base-cyclohexyl, hydroxyl-cyclohexyl, methoxyl group-cyclohexyl, carboxyl-cyclohexyl or hydroxyl-methyl-cyclohexyl base.
16. compound as claimed in claim 1, wherein Y be phenyl, the phenyl that replaced by F, the phenyl that is replaced by C1, the phenyl that is replaced by methoxyl group, the phenyl that is replaced by cyano group, by the dibasic phenyl of F or by the mono-substituted phenyl of ethanoyl.
17. compound as claimed in claim 1, wherein X is methyl-cyclohexyl base or fluoro-2-methyl--cyclohexyl.
18. compound as claimed in claim 15, wherein Y be phenyl, the phenyl that replaced by F, the phenyl that is replaced by C1, the phenyl that is replaced by methoxyl group, the phenyl that is replaced by cyano group, by the dibasic phenyl of F or by the mono-substituted phenyl of ethanoyl.
19. compound as claimed in claim 15, wherein X is methyl-cyclohexyl base or fluoro-2-methyl--cyclohexyl.
20. compound as claimed in claim 18, wherein X is methyl-cyclohexyl base or fluoro-2-methyl--cyclohexyl.
21. compound as claimed in claim 20, wherein Z is hydroxyl-cyclohexyl, methoxyl group-cyclohexyl, carboxyl-cyclohexyl or hydroxyl-methyl-cyclohexyl base.
22. compound as claimed in claim 16, wherein Y is phenyl, 4-fluoro-phenyl, 3-fluoro-phenyl, 4-chloro-phenyl, 4-methoxyl group-phenyl, 4-cyano group-phenyl, 3,4-two fluoro-phenyl or 4-acetylphenyl.
23. compound as claimed in claim 17, wherein X is 4-methyl-cyclohexyl base or 1-fluoro-4-methyl-cyclohexyl base.
24. compound as claimed in claim 1, wherein said compound is selected from:
5-(4-fluoro-phenyl)-3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-thiophene-2-carboxylic acid;
5-(3-fluoro-phenyl)-3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-thiophene-2-carboxylic acid;
5-(4-chloro-phenyl)-3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-thiophene-2-carboxylic acid;
3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-(4-methoxyl group-phenyl)-thiophene-2-carboxylic acid;
5-(4-cyano group-phenyl)-3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-thiophene-2-carboxylic acid;
5-(3,4-two fluoro-phenyl)-3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-thiophene-2-carboxylic acid;
5-(4-ethanoyl-phenyl)-3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-thiophene-2-carboxylic acid;
Or its pharmacy acceptable salt.
25. compound as claimed in claim 24, wherein said compound are 5-(4-fluoro-phenyl)-3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-thiophene-2-carboxylic acid or its pharmacy acceptable salt.
26. compound as claimed in claim 24, wherein said compound are 5-(3-fluoro-phenyl)-3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-thiophene-2-carboxylic acid or its pharmacy acceptable salt.
27. compound as claimed in claim 24, wherein said compound are 5-(4-chloro-phenyl)-3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-thiophene-2-carboxylic acid or its pharmacy acceptable salt.
28. compound as claimed in claim 24, wherein said compound are 3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-(4-methoxyl group-phenyl)-thiophene-2-carboxylic acid or its pharmacy acceptable salt.
29. compound as claimed in claim 24, wherein said compound are 5-(4-cyano group-phenyl)-3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-thiophene-2-carboxylic acid or its pharmacy acceptable salt.
30. compound as claimed in claim 24, wherein said compound are 5-(3,4-two fluoro-phenyl)-3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-thiophene-2-carboxylic acid or its pharmacy acceptable salt.
31. compound as claimed in claim 24, wherein said compound are 5-(4-ethanoyl-phenyl)-3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-thiophene-2-carboxylic acid or its pharmacy acceptable salt.
32. a compound, wherein said compound is selected from:
3-[(4-methyl-cyclohexyl alkyl carbonyl)-(4-oxygen-cyclohexyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
3-[(4-oxyimino-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
3-[(4-methoxyimino-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
3-[(4-methoxyl group-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
3-[(4-carboxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
3-[(3-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid; Or its pharmacy acceptable salt.
33. compound as claimed in claim 32, wherein said compound are 3-[(4-methyl-cyclohexyl alkyl carbonyls)-(4-oxygen-cyclohexyl)-amino]-5-phenyl-thiophene-2-carboxylic acid or its pharmacy acceptable salt.
34. compound as claimed in claim 32, wherein said compound are 3-[(4-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid or its pharmacy acceptable salt.
35. compound as claimed in claim 34, wherein said compound are 3-[(trans-4-hydroxyl-cyclohexyl)-(trans-4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid or its pharmacy acceptable salt.
36. compound as claimed in claim 34, wherein said compound are 3-[(trans-4-methyl-cyclohexyl alkyl carbonyl)-(cis-4-hydroxyl-cyclohexyl)-amino]-5-phenyl-thiophene-2-carboxylic acid or its pharmacy acceptable salt.
37. compound as claimed in claim 32, wherein said compound are 3-[(4-oxyimino-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid or its pharmacy acceptable salt.
38. compound as claimed in claim 32, wherein said compound are 3-[(4-methoxyimino-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid or its pharmacy acceptable salt.
39. compound as claimed in claim 32, wherein said compound are 3-[(4-methoxyl group-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid or its pharmacy acceptable salt.
40. as the compound that claim 32 is gone back, wherein said compound is 3-[(4-carboxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid or its pharmacy acceptable salt.
41. compound as claimed in claim 32, wherein said compound are 3-[(3-hydroxyl-cyclohexyl)-(4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid or its pharmacy acceptable salt.
42. a compound, wherein said following structural formula and the pharmacy acceptable salt thereof of being selected from:
43. compound as claimed in claim 42, wherein said following structural formula and the pharmacy acceptable salt thereof of being selected from:
44. the compound with following formula, or its pharmacy acceptable salt:
Wherein Z is ethyl-piperidyl, sec.-propyl-piperidyl, methyl-oxygen base-piperidyl, ethanoyl-piperidyl, formyl radical-piperidyl, cyano group-piperidyl, methylsulfonyl-piperidyl, oxamoyl base-piperidyl, methylamino formyl radical-piperidyl, benzyl-piperidyl, methoxy-benzyl-oxygen base-piperidyl, the nitrogen heterocyclic heptyl, methyl nitrogen heterocyclic heptyl, oxygen base nitrogen heterocyclic heptyl, hydroxyl-cyclopentyl, hydroxyl-cyclohexyl, methoxyl group-cyclohexyl, carboxyl-cyclohexyl, hydroxyl-methyl-cyclohexyl base, oxygen base-cyclohexyl, oxyimino-cyclohexyl, methoxyimino-cyclohexyl, tetrahydro thiapyran base, 1-oxygen-tetrahydro thiapyran base, or 1,1-dioxy-tetrahydro thiapyran base;
Y is unsubstituted phenyl;
X is the 4-methylcyclohexyl; With
M is 0-1.
45. compound as claimed in claim 44, wherein said compound is selected from:
4-[(2-carboxyl-5-phenyl-thiene-3-yl-)-(trans-4-methyl-cyclohexyl alkyl carbonyl)-amino]-1-methyl-piperidines muriate;
3-[(is trans-4-methyl-cyclohexyl alkyl carbonyl)-(tetrahydrochysene-thiapyran-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
3-[(1,1-dioxy-tetrahydric thiapyran-4-group)-(trans-4-methyl-cyclohexyl alkyl carbonyl)-amino]-5-phenyl-thiophene-2-carboxylic acid;
3-[(is trans-4-methyl-cyclohexyl alkyl carbonyl)-(1-oxygen-tetrahydrochysene-thiapyran-4-yl)-amino]-5-phenyl-thiophene-2-carboxylic acid or its pharmacy acceptable salt.
46. compound as claimed in claim 44; wherein Z is sec.-propyl-piperidyl; methyl-oxygen-piperidyl; ethanoyl-piperidyl; formyl radical-piperidyl; cyano group-piperidyl; methylsulfonyl-piperidyl; oxamoyl base-piperidyl; methylamino formyl radical-piperidyl; benzyl-piperidyl; methoxy-benzyl-oxygen-piperidyl; the nitrogen heterocyclic heptyl; methyl-nitrogen heterocyclic heptyl; oxygen base-nitrogen heterocyclic heptyl; hydroxyl-cyclopentyl; hydroxy-cyclohexyl; methoxyl group-cyclohexyl; carboxyl-cyclohexyl; hydroxyl-methyl-cyclohexyl base; oxygen base-cyclohexyl; oxyimino-cyclohexyl; methoxyimino-cyclohexyl; tetrahydro thiapyran base; 1-oxygen-tetrahydro thiapyran base; or 1,1-dioxy-tetrahydro thiapyran base.
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| US43196402P | 2002-12-10 | 2002-12-10 | |
| US60/431,964 | 2002-12-10 |
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| KR (1) | KR101058696B1 (en) |
| CN (1) | CN100413861C (en) |
| DE (1) | DE60332482D1 (en) |
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- 2003-12-09 DE DE60332482T patent/DE60332482D1/en not_active Expired - Lifetime
- 2003-12-09 ES ES03767343T patent/ES2345438T3/en not_active Expired - Lifetime
- 2003-12-09 PT PT03767343T patent/PT1569929E/en unknown
- 2003-12-09 KR KR1020057010729A patent/KR101058696B1/en not_active IP Right Cessation
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Also Published As
| Publication number | Publication date |
|---|---|
| DE60332482D1 (en) | 2010-06-17 |
| PT1569929E (en) | 2010-06-18 |
| US20080269481A1 (en) | 2008-10-30 |
| CN1795190A (en) | 2006-06-28 |
| US8357718B2 (en) | 2013-01-22 |
| US7402608B2 (en) | 2008-07-22 |
| US8829030B2 (en) | 2014-09-09 |
| KR101058696B1 (en) | 2011-08-22 |
| US20130096106A1 (en) | 2013-04-18 |
| KR20060023109A (en) | 2006-03-13 |
| US20050009804A1 (en) | 2005-01-13 |
| ES2345438T3 (en) | 2010-09-23 |
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